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Sample records for vitro investigative approach

  1. Systematical investigation of in vitro interaction of InP/ZnS quantum dots with human serum albumin by multispectroscopic approach.

    Science.gov (United States)

    Huang, Shan; Qiu, Hangna; Liu, Yi; Huang, Chusheng; Sheng, Jiarong; Cui, Jianguo; Su, Wei; Xiao, Qi

    2016-12-01

    Cadmium-free quantum dots (QDs) have attracted great attention in biological and biomedical applications due to their less content of toxic metals, but their potential toxicity investigations on molecular biology level are rarely involved. Since few studies have addressed whether InP/ZnS QDs could bind and alter the structure and function of human serum albumin (HSA), in vitro interaction between InP/ZnS QDs and HSA was systematically characterized by multispectroscopic approaches. InP/ZnS QDs could quench the intrinsic fluorescence of HSA via static mode. The binding site of InP/ZnS QDs was mainly located at subdomain IIA of HSA. Some thermodynamic parameters suggested that InP/ZnS QDs interacted with HSA mainly through electrostatic interactions. As further revealed by three-dimensional spectrometry, FT-IR spectrometry and circular dichroism technique, InP/ZnS QDs caused more global and local conformational change of HSA than CdSe/ZnS QDs, which illustrated the stronger binding interaction and higher potential toxicity of InP/ZnS QDs on biological function of HSA. Our results offer insights into the in vitro binding mechanism of InP/ZnS QDs with HSA and provide important information for possible toxicity risk of these cadmium-free QDs to human health. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Investigational approaches for mesothelioma

    Directory of Open Access Journals (Sweden)

    Veerle F Surmont

    2011-08-01

    Full Text Available MPM is a rare, aggressive tumour with a poor prognosis. In view of the poor survival benefit from first-line chemotherapy and the lack of subsequent effective treatment options, there is a strong need for the development of more effective treatment approaches for patients with MPM. This review will provide a comprehensive state of the art of new investigational approaches for mesothelioma. In an introductory section, the aetiology, epidemiology, natural history and standard of care treatment for MPM will be discussed. This review provide an update of the major clinical trials that impact mesothelioma treatment, discuss the impact of novel therapeutics and provide perspective on where the clinical research in mesothelioma is moving.The evidence was collected by a systematic analysis of the literature (2000–2011 using the databases Medline (National Library of Medicine, USA, Embase (Elsevier, Netherlands, Cochrane Library (Great Britain, National Guideline Clearinghouse (USA, HTA Database (International Network of Agencies for Health Technology Assessment – INAHTA, NIH database (USA, International Pleural Mesothelioma Program – WHOLIS (WHO Database , with the following keywords and filters: mesothelioma, guidelines, treatment, surgery, chemotherapy, radiotherapy, review, investigational, drugsCurrently different targeted therapies and biologicals are under investigation for MPM. It is important that the molecular biologic research should first focus on mesothelioma-specific pathways and biomarkers in order to have more effective treatment options for this disease. The use of array technology will be certainly an implicit gain in the identification of new potential prognostic or biomarkers or important pathways in the MPM pathogenesis. Probably a central mesothelioma virtual tissue bank may contribute to the ultimate goal to identify druggable targets and to develop personalized treatment for the MPM patients.

  3. Investigation of rabies virus glycoprotein carboxyl terminus as an in vitro predictive tool of neurovirulence. A 3R approach.

    Science.gov (United States)

    Seo, Wonhyo; Prehaud, Christophe; Khan, Zakir; Sabeta, Claude; Lafon, Monique

    In the field of live viral vaccines production, there is an unmet need for in vitro tests complying a 3R approach (Refine, Replace and Reduce the use of animal experimentation) to replace the post-licensing safety tests currently assayed in animals. Here, we performed a pilot study evaluating whether virulence of rabies virus, RABV, can be forecast by an in vitro test of neurite outgrowth. The rationale to use neurite outgrowth as a read-out for this test is based on the salient property of the cytoplasmic domain of the G-protein (Cyto-G) of virulent RABV strains - not of attenuated RABV strains - to stimulate neurite outgrowth in vitro. We observed that neurite elongation triggered by the Cyto-Gs encoded by different RABV field isolates correlate with the distinct virulence scores obtained in a mouse model of experimental rabies. Our results cast the idea that it could be feasible to predict RABV virulence by testing the in vitro property of a RABV strain to promote neurite outgrowth without the use of animal experimentation. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  4. In vitro systems toxicology approach to investigate the effects of repeated cigarette smoke exposure on human buccal and gingival organotypic epithelial tissue cultures

    Science.gov (United States)

    Schlage, Walter K.; Kostadinova, Radina; Xiang, Yang; Sewer, Alain; Majeed, Shoaib; Kuehn, Diana; Frentzel, Stefan; Talikka, Marja; Geertz, Marcel; Mathis, Carole; Ivanov, Nikolai; Hoeng, Julia; Peitsch, Manuel C.

    2014-01-01

    Smoking has been associated with diseases of the lung, pulmonary airways and oral cavity. Cytologic, genomic and transcriptomic changes in oral mucosa correlate with oral pre-neoplasia, cancer and inflammation (e.g. periodontitis). Alteration of smoking-related gene expression changes in oral epithelial cells is similar to that in bronchial and nasal epithelial cells. Using a systems toxicology approach, we have previously assessed the impact of cigarette smoke (CS) seen as perturbations of biological processes in human nasal and bronchial organotypic epithelial culture models. Here, we report our further assessment using in vitro human oral organotypic epithelium models. We exposed the buccal and gingival organotypic epithelial tissue cultures to CS at the air–liquid interface. CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues. Using microarray technology, gene-set analysis and a novel computational modeling approach leveraging causal biological network models, we identified CS impact on xenobiotic metabolism-related pathways accompanied by a more subtle alteration in inflammatory processes. Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset. These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products. PMID:25046638

  5. In vitro systems toxicology approach to investigate the effects of repeated cigarette smoke exposure on human buccal and gingival organotypic epithelial tissue cultures.

    Science.gov (United States)

    Schlage, Walter K; Iskandar, Anita R; Kostadinova, Radina; Xiang, Yang; Sewer, Alain; Majeed, Shoaib; Kuehn, Diana; Frentzel, Stefan; Talikka, Marja; Geertz, Marcel; Mathis, Carole; Ivanov, Nikolai; Hoeng, Julia; Peitsch, Manuel C

    2014-10-01

    Smoking has been associated with diseases of the lung, pulmonary airways and oral cavity. Cytologic, genomic and transcriptomic changes in oral mucosa correlate with oral pre-neoplasia, cancer and inflammation (e.g. periodontitis). Alteration of smoking-related gene expression changes in oral epithelial cells is similar to that in bronchial and nasal epithelial cells. Using a systems toxicology approach, we have previously assessed the impact of cigarette smoke (CS) seen as perturbations of biological processes in human nasal and bronchial organotypic epithelial culture models. Here, we report our further assessment using in vitro human oral organotypic epithelium models. We exposed the buccal and gingival organotypic epithelial tissue cultures to CS at the air-liquid interface. CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues. Using microarray technology, gene-set analysis and a novel computational modeling approach leveraging causal biological network models, we identified CS impact on xenobiotic metabolism-related pathways accompanied by a more subtle alteration in inflammatory processes. Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset. These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products.

  6. Investigation of genetic variability related to the in vitro floral ...

    African Journals Online (AJOL)

    This paper reports on a molecular analysis study conducted on Date palm flowers from the Deglet Nour cultivar to investigate putative genetic variability related to the in vitro floral hermaphrodism induction. Natural male and female as well as hermaphrodite ones that were produced in vitro through the hormonal treatment of ...

  7. Investigating a spatial approach to groundwater quantity ...

    African Journals Online (AJOL)

    2014-12-17

    Dec 17, 2014 ... it takes cognisance of the capture principle, and if the 'precise' directive is only based on a water balance. METHODOLOGY. Overview. The feasibility of a spatial approach, using radius of influence, was investigated from a scientific, a legal, and an institutional perspective. The scientific perspective involved ...

  8. Investigating a spatial approach to groundwater quantity ...

    African Journals Online (AJOL)

    The purpose of this paper is to investigate whether a simple, spatially-based approach to groundwater sustainability using radius of influence should be used to replace the pervasive, yet deprecated, 'natural recharge water balance' volumetric method. Using South Africa as a case study, the radius of influence methodology ...

  9. Parallelogram approach using rat-human In vitro and rat in vivo toxicogenomics predicts acetaminophen-induced hepatotoxicity in humans

    NARCIS (Netherlands)

    Kienhuis, A.S.; Poll, M.C.G. van de; Wortelboer, H.; Herwijnen, M. van; Gottschalk, R.; Jong, C.H.C. de; Boorsma, A.; Paules, R.S.; Kleinjans, J.C.S.; Stierum, R.H.; Delft, J.H.M. van

    2009-01-01

    The frequent use of rodent hepatic in vitro systems in pharmacological and toxicological investigations challenges extrapolation of in vitro results to the situation in vivo and interspecies extrapolation from rodents to humans. The toxicogenomics approach may aid in evaluating relevance of these

  10. Investigation of the cytotoxicity mechanism of silver nanoparticles in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Wei Lina; Zhang Zhixiong; Xi Tingfei [Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Tang Jinglong [Center of Medical Devices, National Institute for the Control of Pharmaceutical and Biological Products, State Food and Drug Administration, Beijing 100050 (China); Chen Yanmei [Department of Advanced Materials and Nanotechnology, College of Engineering, Peking University, Beijing 100871 (China); Zhou Gui, E-mail: xitingfei@tom.co [Precision Medical Devices Department, Shanghai Medical Instrumentation College, Shanghai 200093 (China)

    2010-08-01

    Nowadays, more and more nanotechnology products and nanomaterials are being applied in our lives. Silver nanoparticles (SNPs) are used in infection prevention and treatment due to their antimicrobial activity. However, as a kind of nanomaterial, the toxicology of SNPs has not been completely studied. The mechanism of cytotoxicity of SNPs in vitro to mouse's fibroblast cells (L929) was investigated in this study. As a contrast, silver microparticles (SMPs) were also studied. Propidium iodide (PI) single staining and Annexin-V/PI staining were carried out to unveil the influence of SNPs and SMPs on the cells. A transmission electron microscope (TEM) was used to observe SNPs' distribution in the cells. The results of cell cycle analysis indicated that the cells treated with SNPs were arrested in the G2M phase. Meanwhile, SNPs lead to apoptosis of more cells compared to SMPs at the same dose as a result of apoptosis analysis. Analysis of the cells' ultrastructure showed that SNPs could be phagocytized into the cells while SMPs could not. The mechanism of cytotoxicity of SNPs in vitro to L929 cells may be that SNPs are phagocytized into the cells and they interact with mitochondria or other organelles, even nuclei, which results in cells' apoptosis or necrosis.

  11. In vitro production of alkaloids: Factors, approaches, challenges and prospects

    Science.gov (United States)

    Ahmad, Sayeed; Garg, Madhukar; Tamboli, Ennus Tajuddin; Abdin, M. Z.; Ansari, S. H.

    2013-01-01

    The wide diversity of plant secondary metabolites is largely used for the production of various pharmaceutical compounds. In vitro cell tissue or organ culture has been employed as a possible alternative to produce such industrial compounds. Tissue culture techniques provide continuous, reliable, and renewable source of valuable plant pharmaceuticals and might be used for the large-scale culture of the plant cells from which these secondary metabolites can be extracted. Alkaloids are one of the most important secondary metabolites known to play a vital role in various pharmaceutical applications leading to an increased commercial importance in recent years. The tissue culture techniques may be utilized to improve their production of alkaloids via somaclonal variations and genetic transformations. The focus of this review is toward the application of different tissue culture methods/techniques employed for the in vitro production of alkaloids with a systematic approach to improve their production. PMID:23922453

  12. Systems Approach to Scientific Investigation in Informatics

    Directory of Open Access Journals (Sweden)

    Stanislava Mildeová

    2017-06-01

    Full Text Available The level of informatics development is causally linked to the level of scientific development in the area. Successful scientific work is based not only on the investigative style of thinking, but it also means to think in context and to see the whole setting of the solved research problem – to be able to apply system approaches. System approaches include System Dynamics; a community practicing system dynamics has become the strongest school of system approach with many members of the academic environment as well as from real life practice. The aim of the article is to show the possibilities of system dynamics models as appropriate tools to support system approach to scientific research in informatics. The multidisciplinary and quantitative concepts that are typical of System Dynamics are, according to the authors, suitable for systemic solution of the complex problems of current IT practice. Conducted research and examining the principles of system dynamics including the stages of creation of its models confirm these author's arguments.

  13. An in vitro investigation of the erosive potential of smoothies.

    Science.gov (United States)

    Blacker, S M; Chadwick, R G

    2013-02-01

    Recent health promotion campaigns have encouraged the public to consume at least five portions of fruit and vegetables per day. Many see consuming fruit smoothies as a way of achieving this. To ascertain the potential or otherwise for fruit smoothies to bring about dental erosion. Laboratory study.Method This was an in vitro investigation in which five varieties of shop bought fruit smoothies, including a 'thickie' were investigated, with respect to their initial pH, titratable acidity and effect upon exposure to the surface microhardness and profile of extracted human teeth. In addition their performance was compared to negative (Volvic(™) water) and positive (orange juice) control drinks as well as a homemade smoothie, based upon the recipe of one of the commercially bought drinks, from which ingredient omissions were made. The majority of the drinks investigated had a baseline pH below the critical pH of enamel (5.5) and required comparable volumes of 0.1M NaOH to raise their pH to neutrality as the positive control. Only two drinks (Volvic(™) still mineral water, the negative control, and the yoghurt, vanilla bean and honey 'thickie') displayed a higher pH, though to neutralise the thickie, a lesser quantity of alkali addition was required. The immersion of the tooth samples in the drinks brought about reductions in their surface hardness (expressed as a percentage change of median hardness) but these were only significant (p benefits of their consumption, their consumption should be confined to mealtimes.

  14. In vitro tests and ethnopharmacological investigations: wound healing as an example.

    Science.gov (United States)

    Houghton, P J; Hylands, P J; Mensah, A Y; Hensel, A; Deters, A M

    2005-08-22

    In vitro tests are now widely employed in ethnopharmacological research because of ethical reasons and their usefulness in bioactive-guided fractionation and determination of active compounds. For many disease conditions, a variety of in vitro tests can now be employed as the biochemical mechanisms underlying disease and healing processes are understood. Approaches to the in vitro investigations of wound healing processes are exemplified by studies on extracts of Buddleja species and three Ghanaian species Spathodea campanulata, Commelina diffusa and Secamone afzelii. Most studies have been carried out on Buddleja officinalis or Buddleja globosa. The extracts have been shown to have anti-inflammatory and antioxidant properties due to flavonoids, triterpenoids, diterpenoids and caffeic acid derivatives. There appears to a slight effect on proliferation of fibroblasts at lower concentrations, but this was not significant, and higher concentrations appeared to be cytotoxic. Novel findings are the ability of Buddleja globosa leaf extracts to induce differentiation in keratinocytes and to alter the profile of proteins produced by cultured fibroblasts. Extracts also had some effect on lattice contraction. The three Ghanaian species examined show a mixture of antimicrobial and antioxidant activities. The evolution over recent years of tests for wound healing, from in vivo tests to cell-based systems and chemical reactions and on to investigations into effects on secondary messengers and protein expression, is described.

  15. A Computational Approach to Understand In Vitro Alveolar Morphogenesis

    Science.gov (United States)

    Kim, Sean H. J.; Yu, Wei; Mostov, Keith; Matthay, Michael A.; Hunt, C. Anthony

    2009-01-01

    Primary human alveolar type II (AT II) epithelial cells maintained in Matrigel cultures form alveolar-like cysts (ALCs) using a cytogenesis mechanism that is different from that of other studied epithelial cell types: neither proliferation nor death is involved. During ALC formation, AT II cells engage simultaneously in fundamentally different, but not fully characterized activities. Mechanisms enabling these activities and the roles they play during different process stages are virtually unknown. Identifying, characterizing, and understanding the activities and mechanisms are essential to achieving deeper insight into this fundamental feature of morphogenesis. That deeper insight is needed to answer important questions. When and how does an AT cell choose to switch from one activity to another? Why does it choose one action rather than another? We report obtaining plausible answers using a rigorous, multi-attribute modeling and simulation approach that leveraged earlier efforts by using new, agent and object-oriented capabilities. We discovered a set of cell-level operating principles that enabled in silico cells to self-organize and generate systemic cystogenesis phenomena that are quantitatively indistinguishable from those observed in vitro. Success required that the cell components be quasi-autonomous. As simulation time advances, each in silico cell autonomously updates its environment information to reclassify its condition. It then uses the axiomatic operating principles to execute just one action for each possible condition. The quasi-autonomous actions of individual in silico cells were sufficient for developing stable cyst-like structures. The results strengthen in silico to in vitro mappings at three levels: mechanisms, behaviors, and operating principles, thereby achieving a degree of validation and enabling answering the questions posed. We suggest that the in silico operating principles presented may have a biological counterpart and that a

  16. A simulation framework to investigate in vitro viral infection dynamics

    NARCIS (Netherlands)

    Bankhead, A.; Mancini, E.; Sims, A.C.; Baric, R.S.; McWeeney, S.; Sloot, P.M.A.

    2011-01-01

    Virus infection is a complex biological phenomenon for which in vitro experiments provide a uniquely concise view where data is often obtained from a single population of cells, under controlled environmental conditions. Nonetheless, data interpretation and real understanding of viral dynamics is

  17. New approach to Ice Complex investigation

    Science.gov (United States)

    Tumskoy, V. E.

    2015-12-01

    Ice Complex (IC) is the most well-known type of syncryogenic permafrost deposit. ICs are widely distributed in the northern-eastern part of Russia (Siberia) and in Alaska. These deposits are an important source of paleo-climatic information, because they contain well-preserved remains of flora and fauna typical for the time of preservation. The isotope signature of ice preserved in an IC can help to reconstruct specific features of the freezing period and freezing conditions. However, the best methods to use to determine the origin and age of ICs and what constitutes typical IC morphological structure are still being discussed. Conventionally, syncryogenic permafrost deposits of Pleistocene age, consisting of an ice-wedge net, are considered ICs. In our multi-year study, we applied a new approach, defining ICs using cryolithological criteria. Our results allow us to conclude that the initial composition of an IC is determined by the sediments (silt, sand, gravel) and processes (fluvial, slope, aeolian) involved in IC genesis. Further alteration of IC composition occurs via cryogenic processes during glacial epochs and partial/complete thawing during inter-glacial epochs; usually such alteration leads to an increase in the fine-grain fraction of IC sediments and the formation of separate IC layers, which should be considered as separate stratigraphic units. Only the uppermost unit of an IC should be called an edoma ice complex (EIC); this unit creates specific relief features, for which EICs are known, in Siberia and Yakutia. The most important task to which attention should be paid in further investigations is the development of criteria that will allow reconstructive modeling of the processes and conditions that existed in the past, processes and conditions which determined initial IC composition and alteration of properties over time.

  18. Characteristic MR spectroscopy in fucosidosis: in vitro investigation

    Energy Technology Data Exchange (ETDEWEB)

    Mamourian, Alex C.; Chawla, Sanjeev; Poptani, Harish [Hospital of the University of Pennsylvania, Department of Radiology, Philadelphia, PA (United States); Hopkin, Jeremy R. [Barrow Neurological Institute, Department of Neuroradiology, Phoenix, AZ (United States)

    2010-08-15

    Fucosidosis is a rare lysosomal storage disorder that results in the deposition of the sugar fucose within various organs, including the central nervous system. Neuroimaging abnormalities on MR, specifically T2 shortening in the basal ganglia, have been reported as suggestive of fucosidosis. A more recent report of MR spectroscopy (MRS) of one patient provided evidence that MRS is specific for fucosidosis. We present another confirmed case with nearly identical MR spectroscopic findings along with in vitro data that support the contention that MR spectroscopy, in the setting of typical clinical and imaging features, is characteristic for this rare disorder. (orig.)

  19. Real-time closed-loop electrophysiology: towards new frontiers in in vitro investigations in the neurosciences.

    Science.gov (United States)

    Arsiero, M; Lüscher, H-R; Giugliano, M

    2007-11-01

    Reflected at any level of organization of the central nervous system, most of the processes ranging from ion channels to neuronal networks occur in a closed loop, where the input to the system depends on its output. In contrast, most in vitro preparations and experimental protocols operate autonomously, and do not depend on the output of the studied system. Thanks to the progress in digital signal processing and real-time computing, it is now possible to artificially close the loop and investigate biophysical processes and mechanisms under increased realism. In this contribution, we review some of the most relevant examples of a new trend in in vitro electrophysiology, ranging from the use of dynamic-clamp to multi-electrode distributed feedback stimulation. We are convinced these represents the beginning of new frontiers for the in vitro investigation of the brain, promising to open the still existing borders between theoretical and experimental approaches while taking advantage of cutting edge technologies.

  20. An investigation of fake fingerprint detection approaches

    Science.gov (United States)

    Ahmad, Asraful Syifaa'; Hassan, Rohayanti; Othman, Razib M.

    2017-10-01

    The most reliable biometrics technology, fingerprint recognition is widely used in terms of security due to its permanence and uniqueness. However, it is also vulnerable to the certain type of attacks including presenting fake fingerprints to the sensor which requires the development of new and efficient protection measures. Particularly, the aim is to identify the most recent literature related to the fake fingerprint recognition and only focus on software-based approaches. A systematic review is performed by analyzing 146 primary studies from the gross collection of 34 research papers to determine the taxonomy, approaches, online public databases, and limitations of the fake fingerprint. Fourteen software-based approaches have been briefly described, four limitations of fake fingerprint image were revealed and two known fake fingerprint databases were addressed briefly in this review. Therefore this work provides an overview of an insight into the current understanding of fake fingerprint recognition besides identifying future research possibilities.

  1. In vitro antiplasmodial investigation of medicinal plants from El Salvador.

    Science.gov (United States)

    Köhler, Inga; Jenett-Siems, Kristina; Siems, Karsten; Hernández, Marco Antonio; Ibarra, Ricardo A; Berendsohn, Walter G; Bienzle, Ulrich; Eich, Eckart

    2002-01-01

    In vitro antiplasmodial activities of extracts from Albizia saman, Fabaceae, Calea tenuifolia (C. zacatechichi), Asteraceae, Hymenaea courbaril, Fabaceae, Jatropha curcas, Euphorbiaceae, Momordica charantia, Cucurbitaceae, and Moringa oleifera, Moringaceae were evaluated. From the lipophilic extract of C tenuifolia five active flavones were obtained. 4',5-Dihydroxy-7-methoxyflavone [genkwanin] and 5-hydroxy-4',7-dimethoxyflavone [apigenin 4',7-dimethylether] exhibited the strongest antiplasmodial activity against a chloroquine-sensitive strain (poW) and a chloroquine-resistant strain (Dd2) of Plasmodium falciparum [IC50 values: 17.1-28.5 microM). Furthermore octadeca-9,12-dienoic acid [linoleic acid] [IC50] values of 21.8 microM (poW) and 31.1 microM (Dd2)] and octadeca-9,12,15-trienoic acid (alpha-linolenic acid) were isolated.

  2. In vitro investigation of Debaryomyces hansenii strains for potential probiotic properties

    DEFF Research Database (Denmark)

    Ochangco, Honeylet Sabas; Gamero, Amparo; Smith, Ida Mosbech

    2016-01-01

    In this study, 23 Debaryomyces hansenii strains, isolated from cheese and fish gut, were investigated in vitro for potential probiotic properties i.e. (1) survival under in vitro GI (gastrointestinal) conditions with different oxygen levels, (2) adhesion to Caco-2 intestinal epithelial cells and ...

  3. A European Approach to Clinical Investigator Training

    Directory of Open Access Journals (Sweden)

    Jean-Marie eBoeynaems

    2013-09-01

    Full Text Available A better education and training of clinical investigators and their teams is one of the factors that could foster the development of clinical research in Europe, a key objective of the Innovative Medicines Initiative (IMI. PharmaTrain (an IMI programme on training in medicines development and ECRIN (European Clinical Research Infrastructures Network have joined forces to address this issue. An advisory group composed of representatives of universities, pharmaceutical companies and other organisations met four times between June 2011 and July 2012. This resulted in a position paper proposing a strategy to improve and harmonize clinical investigator training in Europe, and including a detailed syllabus and list of learning outcomes. Major recommendations are the establishment of minimal and mutually recognized certification requirement for investigators throughout the EU and the creation of a European platform to provide a suitable course and examination infrastructure.

  4. A novel approach for in vitro meat production.

    Science.gov (United States)

    Pandurangan, Muthuraman; Kim, Doo Hwan

    2015-07-01

    The present review describes the possibility of in vitro meat production with the help of advanced co-culturing methods. In vitro meat production method could be a possible alternative for the conventional meat production. Originally, the research on in vitro meat production was initiated by the National Aeronautics and Space Administration (NASA) for space voyages. The required key qualities for accepting in vitro meat for consumption would be good efficiency ratio, increased protein synthesis rate in skeletal muscles, and mimicking the conventional meat qualities. In vitro culturing of meat is possible with the use of skeletal muscle tissue engineering, stem cell, cell co-culture, and tissue culture methods. Co-culture of myoblast and fibroblast is believed as one of the major techniques for in vitro meat production. In our lab, we have co-cultured myoblast and fibroblast. We believe that a billion pounds of in vitro meat could be produced from one animal for consumption. However, we require a great deal of research on in vitro meat production.

  5. Salinity Stress Tolerance Of Camelina Investigated In Vitro

    Directory of Open Access Journals (Sweden)

    Khalid H.

    2015-12-01

    Full Text Available The ability of Camelina sativa to withstand salinity stress in vitro by adding NaCl (0, 25, 50, 75, 100, 125, 150, 175, 200mM in Murashige and Skoog basal medium was studied. Performance of the plants was measured in terms of various growth parameters and physiological and biochemical tests performed on fully grown plants. The germination capacity, cotyledon unfolding and first true leaf emergence was reduced by 30.6, 17.3, and 28.8%, respectively in 200mM salt treatment with respect to control. The plant height, relative water content, and plant water content were decreased by 85.4, 10.8, and 9.8%, respectively, in stressed plants with respect to control. A decrease in chlorophyll a and b and total chlorophyll contents (by 81.3%, as well as of protein content was registered. Electrical conductivity increased by 52.8% in stressed plants over control, as expected. Other stress indicators like guiacol peroxidase activity and malondialdehyde also increased with respect to control. At salt concentrations lower than 200mM, no clear cut retardation effects were seen. Thus, the present study opens up the scope of further assessment of survivability of camelina in salt contaminated soils.

  6. Middle Ear Prosthesis with Bactericidal Efficacy—In Vitro Investigation

    Directory of Open Access Journals (Sweden)

    Magdalena Ziąbka

    2017-10-01

    Full Text Available Materials used in ossicular replacement prostheses must possess appropriate biological properties, such as biocompatibility, stability, no cytotoxicity. Due to the risk of infection (otitis media and chronic otitis media, it is desirable to use an antibacterial agent for illness prevention during the ossicular reconstruction. The goal of this work was to observe biological properties of a new composite prosthesis made of ABS containing silver nanoparticles (AgNPs 45T. Samples for biological tests and then a prototype of middle ear prosthesis were prepared using injection moulding and extrusion techniques. In vitro experiments were carried out to assess bactericidal efficacy against Staphylococcus aureus and Pseudomona aeruginosa standard strains, cell proliferation, viability and cytotoxicity, using Hs680.Tr. fibroblast cells. Surface parameters of the samples were evaluated, including roughness and wettability. The silver ions were continually released from the polymer in aqueous solution. The silver ions release was measured as increasing with time and concentration of the silver nanoparticles in the polymer matrix. No cytotoxicity effect was observed, while bactericidal efficacy was noticed for silver nanoparticles. The roughness studies showed an increase in roughness for the samples with silver nanoparticles. All polymer and composite materials containing silver nanoparticles showed hydrophilic properties. The composites were found to release silver ions at a concentration level capable of rendering the antimicrobial efficacy even with the lowest concentration of silver nanoparticles in the material. Our results demonstrate that middle ear prosthesis made of polymer and silver nanoparticles may eliminate bacteria during inflammation in the middle ear.

  7. Middle Ear Prosthesis with Bactericidal Efficacy-In Vitro Investigation.

    Science.gov (United States)

    Ziąbka, Magdalena; Dziadek, Michał; Menaszek, Elżbieta; Banasiuk, Rafał; Królicka, Aleksandra

    2017-10-10

    Materials used in ossicular replacement prostheses must possess appropriate biological properties, such as biocompatibility, stability, no cytotoxicity. Due to the risk of infection (otitis media and chronic otitis media), it is desirable to use an antibacterial agent for illness prevention during the ossicular reconstruction. The goal of this work was to observe biological properties of a new composite prosthesis made of ABS containing silver nanoparticles (AgNPs 45T). Samples for biological tests and then a prototype of middle ear prosthesis were prepared using injection moulding and extrusion techniques. In vitro experiments were carried out to assess bactericidal efficacy against Staphylococcus aureus and Pseudomona aeruginosa standard strains, cell proliferation, viability and cytotoxicity, using Hs680.Tr. fibroblast cells. Surface parameters of the samples were evaluated, including roughness and wettability. The silver ions were continually released from the polymer in aqueous solution. The silver ions release was measured as increasing with time and concentration of the silver nanoparticles in the polymer matrix. No cytotoxicity effect was observed, while bactericidal efficacy was noticed for silver nanoparticles. The roughness studies showed an increase in roughness for the samples with silver nanoparticles. All polymer and composite materials containing silver nanoparticles showed hydrophilic properties. The composites were found to release silver ions at a concentration level capable of rendering the antimicrobial efficacy even with the lowest concentration of silver nanoparticles in the material. Our results demonstrate that middle ear prosthesis made of polymer and silver nanoparticles may eliminate bacteria during inflammation in the middle ear.

  8. Computational investigation of epithelial cell dynamic phenotype in vitro

    Directory of Open Access Journals (Sweden)

    Debnath Jayanta

    2009-05-01

    Full Text Available Abstract Background When grown in three-dimensional (3D cultures, epithelial cells typically form cystic organoids that recapitulate cardinal features of in vivo epithelial structures. Characterizing essential cell actions and their roles, which constitute the system's dynamic phenotype, is critical to gaining deeper insight into the cystogenesis phenomena. Methods Starting with an earlier in silico epithelial analogue (ISEA1 that validated for several Madin-Darby canine kidney (MDCK epithelial cell culture attributes, we built a revised analogue (ISEA2 to increase overlap between analogue and cell culture traits. Both analogues used agent-based, discrete event methods. A set of axioms determined ISEA behaviors; together, they specified the analogue's operating principles. A new experimentation framework enabled tracking relative axiom use and roles during simulated cystogenesis along with establishment of the consequences of their disruption. Results ISEA2 consistently produced convex cystic structures in a simulated embedded culture. Axiom use measures provided detailed descriptions of the analogue's dynamic phenotype. Dysregulating key cell death and division axioms led to disorganized structures. Adhering to either axiom less than 80% of the time caused ISEA1 to form easily identified morphological changes. ISEA2 was more robust to identical dysregulation. Both dysregulated analogues exhibited characteristics that resembled those associated with an in vitro model of early glandular epithelial cancer. Conclusion We documented the causal chains of events, and their relative roles, responsible for simulated cystogenesis. The results stand as an early hypothesis–a theory–of how individual MDCK cell actions give rise to consistently roundish, cystic organoids.

  9. Endocrine Disruption and In Vitro Ecotoxicology: Recent Advances and Approaches.

    Science.gov (United States)

    Wagner, Martin; Kienle, Cornelia; Vermeirssen, Etiënne L M; Oehlmann, Jörg

    Endocrine-disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling. Omnipresent in the environment, they can cause adverse effects in a wide range of wildlife. Accordingly, Endocrine Disruption is one focal area of ecotoxicology. Because EDCs induce complex response patterns in vivo via a wide range of mechanisms of action, in vitro techniques have been developed to reduce and understand endocrine toxicity. In this review we revisit the evidence for endocrine disruption in diverse species and the underlying molecular mechanisms. Based on this, we examine the battery of in vitro bioassays currently in use in ecotoxicological research and discuss the following key questions. Why do we use in vitro techniques? What endpoints are we looking at? Which applications are we using in vitro bioassays for? How can we put in vitro data into a broader context? And finally, what is the practical relevance of in vitro data? In critically examining these questions, we review the current state-of-the-art of in vitro (eco)toxicology, highlight important limitations and challenges, and discuss emerging trends and future research needs.

  10. Cholesterol assimilation by Lactobacillus probiotic bacteria: an in vitro investigation.

    Science.gov (United States)

    Tomaro-Duchesneau, Catherine; Jones, Mitchell L; Shah, Divya; Jain, Poonam; Saha, Shyamali; Prakash, Satya

    2014-01-01

    Excess cholesterol is associated with cardiovascular diseases (CVD), an important cause of mortality worldwide. Current CVD therapeutic measures, lifestyle and dietary interventions, and pharmaceutical agents for regulating cholesterol levels are inadequate. Probiotic bacteria have demonstrated potential to lower cholesterol levels by different mechanisms, including bile salt hydrolase activity, production of compounds that inhibit enzymes such as 3-hydroxy-3-methylglutaryl coenzyme A, and cholesterol assimilation. This work investigates 11 Lactobacillus strains for cholesterol assimilation. Probiotic strains for investigation were selected from the literature: Lactobacillus reuteri NCIMB 11951, L. reuteri NCIMB 701359, L. reuteri NCIMB 702655, L. reuteri NCIMB 701089, L. reuteri NCIMB 702656, Lactobacillus fermentum NCIMB 5221, L. fermentum NCIMB 8829, L. fermentum NCIMB 2797, Lactobacillus rhamnosus ATCC 53103 GG, Lactobacillus acidophilus ATCC 314, and Lactobacillus plantarum ATCC 14917. Cholesterol assimilation was investigated in culture media and under simulated intestinal conditions. The best cholesterol assimilator was L. plantarum ATCC 14917 (15.18±0.55 mg/10(10) cfu) in MRS broth. L. reuteri NCIMB 701089 assimilated over 67% (2254.70±63.33 mg/10(10) cfu) of cholesterol, the most of all the strains, under intestinal conditions. This work demonstrates that probiotic bacteria can assimilate cholesterol under intestinal conditions, with L. reuteri NCIMB 701089 showing great potential as a CVD therapeutic.

  11. Bio-active nanoemulsions enriched with gold nanoparticle, marigold extracts and lipoic acid: In vitro investigations.

    Science.gov (United States)

    Guler, Emine; Barlas, F Baris; Yavuz, Murat; Demir, Bilal; Gumus, Z Pinar; Baspinar, Yucel; Coskunol, Hakan; Timur, Suna

    2014-09-01

    A novel and efficient approach for the preparation of enriched herbal formulations was described and their potential applications including wound healing and antioxidant activity (cell based and cell free) were investigated via in vitro cell culture studies. Nigella sativa oil was enriched with Calendula officinalis extract and lipoic acid capped gold nanoparticles (AuNP-LA) using nanoemulsion systems. The combination of these bio-active compounds was used to design oil in water (O/W) and water in oil (W/O) emulsions. The resulted emulsions were characterized by particle size measurements. The phenolic content of each nanoemulsion was examined by using both colorimetric assay and chromatographic analyses. Two different methods containing cell free chemical assay (1-diphenyl-2-picrylhydrazyl method) and cell based antioxidant activity test were used to evaluate the antioxidant capacities. In order to investigate the bio-activities of the herbal formulations, in vitro cell culture experiments, including cytotoxicity, scratch assay, antioxidant activity and cell proliferation were carried out using Vero cell line as a model cell line. Furthermore, to monitor localization of the nanoemulsions after application of the cell culture, the cell images were monitored via fluorescence microscope after FITC labeling. All data confirmed that the enriched N. sativa formulations exhibited better antioxidant and wound healing activity than N. sativa emulsion without any enrichment. In conclusion, the incorporation of AuNP-LA and C. officinalis extract into the N. sativa emulsions significantly increased the bio-activities. The present work may support further studies about using the other bio-active agents for the enrichment of herbal preparations to strengthen their activities. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Alternative approaches to ovarian stimulation for in vitro fertilization

    NARCIS (Netherlands)

    D. de Jong (Diederick)

    2003-01-01

    markdownabstract__Abstract__ In vitro fertilization (IVF), literally meaning ‘fertilization in glass’, is characterized by co-culture of aspired oocytes with spermatozoa and subsequent transfer of the embryo(s) into the uterine cavity. The original indication for this treatment was

  13. Expert consensus on an in vitro approach to assess ...

    Science.gov (United States)

    Report from an international workshop with the goal of reviewing the state-of-the-science and determine the technical needs to develop an in vitro system that will reduce and eventually replace the use of animals for evaluating the potential inhalation toxicity of nanomaterials (NMs) in a regulatory setting. Workshop was co-organized in February 2015 by the PETA International Science Consortium Ltd. with the National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods an international workshop that was attended by representatives from industry, government, academia, and non-governmental organizations with expertise in in vivo and in vitro lung systems, respiratory toxicology, inhalation particle dosimetry, nanotoxicology, and hazard and human health risk analysis. This report provides an overview of the presentations, discussions, and recommendations of the participants on the design of an in vitro system for the prediction of pulmonary fibrosis. The workshop participants identified multi-walled carbon nanotubes (MWCNTs), which have been shown to induce fibrosis in animal experiments and represent an important commercial nanomaterial class, as representative pro-fibrogenic NMs to use for the development of an in vitro test system. Recommendations were made for designing a system using lung relevant cells co-cultured at the air-liquid interface to assess the pro-fibrogenic potential of aerosolized MWCNTs, while consider

  14. A CONCEPTUAL REVIEW ON LINGUISTIC APPROACHES OF IDENTITY INVESTIGATION

    Directory of Open Access Journals (Sweden)

    Meinarni Susilowati

    2014-12-01

    Full Text Available Identity has been a blossoming issue in different fields. The intensity of investigating identity has stimulated the diverse methods and approaches to study identity from different angles. This paper discusses the how identity can be investigated from three different linguistic approaches, sociolinguistics, discourse analysis, and sociocultural linguistics approach. The practicality of these three approaches is explored to detect the nature of identity which is fluid, multiple, fragmented, socially, culturally, historically, religiously, and politically constructed and emerges within interactions. More space, however, is invested for elaborating the five principles of sociocultural linguistic approach due to its flexibility and multidimension of the approach. Empirical data is provided for proving its practicality for identity investigation. Further areas of investigation is given at the last part of the paper.

  15. Can magnetic resonance spectroscopy differentiate malignant and benign causes of lymphadenopathy? An in-vitro approach

    National Research Council Canada - National Science Library

    Lionel Buré; Louis-Martin Boucher; Miriam Blumenkrantz; Stefan Schob; Pierre Lafaye de Micheaux; Caroline Reinhold; Benoit Gallix

    2017-01-01

    .... The goal of our research was to assess whether magnetic resonance (MR) spectroscopy contains the necessary information to allow differentiation of benign from malignant lymph nodes in an in-vitro approach using a modern pattern recognition method...

  16. Estrogenicity of halogenated bisphenol A: in vitro and in silico investigations.

    Science.gov (United States)

    Zhang, Jie; Li, Tiezhu; Wang, Tuoyi; Yuan, Cuiping; Zhong, Shuning; Guan, Tianzhu; Li, Zhuolin; Wang, Yongzhi; Yu, Hansong; Luo, Quan; Wang, Yongjun; Zhang, Tiehua

    2017-11-20

    The binding interactions of bisphenol A (BPA) and its halogenated derivatives (halogenated BPAs) to human estrogen receptor α ligand binding domain (hERα-LBD) was investigated using a combined in vitro and in silico approach. First, the recombinant hERα-LBD was prepared as a soluble protein in Escherichia coli BL21(DE3)pLysS. A native fluorescent phytoestrogen, coumestrol, was employed as tracer for the fluorescence polarization assay. The results of the in vitro binding assay showed that bisphenol compounds could bind to hERα-LBD as the affinity ligands. All the tested halogenated BPAs exhibited weaker receptor binding than BPA, which might be explained by the steric effect of substituents. Molecular docking studies elucidated that the halogenated BPAs adopted different conformations in the flexible hydrophobic ligand binding pocket (LBP), which is mainly dependent on their distinct halogenation patterns. The compounds with halogen substituents on the phenolic rings and on the bridging alkyl moiety acted as agonists and antagonists for hERα, respectively. Interestingly, all the compounds in the agonist conformation of hERα formed a hydrogen bond with His524, while the compounds in the antagonist conformation formed a hydrogen bond with Thr347. These docking results suggested a pivotal role of His524/Thr347 in maintaining the hERα structure in the biologically active agonist/antagonist conformation. Comparison of the calculated binding energies vs. experimental binding affinities yielded a good correlation, which might be applicable for the structure-based design of novel bisphenol compounds with reduced toxicities and for environmental risk assessment. In addition, based on hERα-LBD as a recognition element, the proposed fluorescence polarization assay may offer an alternative to chromatographic techniques for the multi-residue determination of bisphenol compounds.

  17. Investigation of In-Vitro Biological Behavior and Pro-Angiogenic ...

    African Journals Online (AJOL)

    Prof. Ogunji

    Investigation of In-Vitro Biological Behavior and Pro-Angiogenic. Potential of Baicalein as ... Current research is tackling the problem of vascularization with four distinct strategies, all of which have demonstrated ... To overcome this problem a lot of research effort has gone into the search for hypoxia mimics. (Xia et al., 2009, ...

  18. Tissue Engineering Approaches in the Design of Healthy and Pathological In Vitro Tissue Models

    Science.gov (United States)

    Caddeo, Silvia; Boffito, Monica; Sartori, Susanna

    2017-01-01

    In the tissue engineering (TE) paradigm, engineering and life sciences tools are combined to develop bioartificial substitutes for organs and tissues, which can in turn be applied in regenerative medicine, pharmaceutical, diagnostic, and basic research to elucidate fundamental aspects of cell functions in vivo or to identify mechanisms involved in aging processes and disease onset and progression. The complex three-dimensional (3D) microenvironment in which cells are organized in vivo allows the interaction between different cell types and between cells and the extracellular matrix, the composition of which varies as a function of the tissue, the degree of maturation, and health conditions. In this context, 3D in vitro models can more realistically reproduce a tissue or organ than two-dimensional (2D) models. Moreover, they can overcome the limitations of animal models and reduce the need for in vivo tests, according to the “3Rs” guiding principles for a more ethical research. The design of 3D engineered tissue models is currently in its development stage, showing high potential in overcoming the limitations of already available models. However, many issues are still opened, concerning the identification of the optimal scaffold-forming materials, cell source and biofabrication technology, and the best cell culture conditions (biochemical and physical cues) to finely replicate the native tissue and the surrounding environment. In the near future, 3D tissue-engineered models are expected to become useful tools in the preliminary testing and screening of drugs and therapies and in the investigation of the molecular mechanisms underpinning disease onset and progression. In this review, the application of TE principles to the design of in vitro 3D models will be surveyed, with a focus on the strengths and weaknesses of this emerging approach. In addition, a brief overview on the development of in vitro models of healthy and pathological bone, heart, pancreas, and

  19. Protein Engineering by Combined Computational and In Vitro Evolution Approaches.

    Science.gov (United States)

    Rosenfeld, Lior; Heyne, Michael; Shifman, Julia M; Papo, Niv

    2016-05-01

    Two alternative strategies are commonly used to study protein-protein interactions (PPIs) and to engineer protein-based inhibitors. In one approach, binders are selected experimentally from combinatorial libraries of protein mutants that are displayed on a cell surface. In the other approach, computational modeling is used to explore an astronomically large number of protein sequences to select a small number of sequences for experimental testing. While both approaches have some limitations, their combination produces superior results in various protein engineering applications. Such applications include the design of novel binders and inhibitors, the enhancement of affinity and specificity, and the mapping of binding epitopes. The combination of these approaches also aids in the understanding of the specificity profiles of various PPIs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Magnetic stent hyperthermia for esophageal cancer: an in vitro investigation in the ECA-109 cell line.

    Science.gov (United States)

    Liu, Jia-Yi; Zhao, Ling-Yun; Wang, Yu-Ying; Li, Dan-Ye; Tao, Dan; Li, Li-Ya; Tang, Jin-Tian

    2012-03-01

    Magnetic stent hyperthermia (MSH) is a novel approach for targeted thermotherapy for esophageal cancer, which is based on the mechanism that inductive heat can be generated by the esophageal stent upon exposure under an alternative magnetic field (AMF). A positive effect of MSH on esophageal cancer has been demonstrated, however, there is no study on the in vitro effects of heating treatment or of the effects of AMF exposure on human esophageal cancer cells. This study aimed to investigate the effect of MSH and of AMF exposure in esophageal cancer cells. Inductive heating characteristics of esophageal stents were assessed by exposing the stents under AMF. A rather rapid temperature rise of the Ni-Ti stent when subjected to AMF exposure was observed and the desired hyperthermic temperature could be controlled by adjusting the field parameter of the AMF. Human esophageal squamous carcinoma (ESCC) ECA-109 cells were divided into four groups: the control group, the water-bath heating group, the MSH group and the AMF exposure group. Hyperthermic temperatures were 43, 48 and 53˚C and the treatment time was in the range of 5-30 min. The MTT assay, apoptotic analysis and TUNEL staining were applied in the current investigation. Exposure of ECA-109 cells under AMF with a field intensity of 50 to 110 kA/m had negligible effect on cell viability, cell necrosis and apoptosis. Hyperthermia had a remarkable inhibitory effect on the cell viability and the effect was dependent on the thermal dose (temperature and time). The optimal thermal dose of MSH for ECA-109 cells was 48˚C for 20-30 min. The study also elucidated that there was a difference in the effects on cell necrosis and apoptosis between the heating mode of water bath and MSH. The data suggest that MSH may have clinical significance for esophageal cancer treatment.

  1. Investigating design: A comparison of manifest and latent approaches

    DEFF Research Database (Denmark)

    Cash, Philip; Snider, Chris

    2014-01-01

    them together and explicitly investigating their complementarity in the design domain. This is realised using an example artificial observation study. In addition to discussing underlying relationships between the approaches, this paper identifies key opportunities for improving design research methods......This paper contributes to the on-going focus on improving design research methods, by exploring and synthesising two key interrelated research approaches e manifest and latent. These approaches are widely used individually in design research, however, this paper represents the first work bringing...... by more explicitly combining both manifest and latent elements. Finally, a number of combinatory approaches are proposed based on a conceptual framework. © 2014 Elsevier Ltd. All rights reserved....

  2. Zinc oxide nanoparticles as a novel anticancer approach; in vitro and in vivo evidence.

    Science.gov (United States)

    Hassan, Hesham Fathy Hassan; Mansour, Ahmed Mohamed; Abo-Youssef, Amira Morad Hussein; Elsadek, Bakheet E M; Messiha, Basim Anwar Shehata

    2017-02-01

    Currently, the outcomes of conventional chemotherapeutic approaches are unsatisfactory. Clinical application of nanoparticles seems promising. We aim to evaluate the possible antitumor activity of zinc oxide nanoparticles (ZnONPs) as a chemotherapeutic approach in in vitro and in vivo experimental models. An in vitro study was performed on three different cell lines, namely human hepatocellular carcinoma (HEPG2), human prostate cancer (PC3), and none-small cell lung cancer (A549) cell lines. An in vivo study using diethylnitrosamine (DENA)-induced HCC in adult male Wistar rats was conducted to investigate the potential antitumor activity of ZnONPs in HCC and the possible underlying mechanisms. Hepatocellular carcinoma (HCC) was induced by oral administration of DENA given in drinking water (100 mg/L) for 8 weeks. Rats were allocated into four groups, namely a control group, an HCC control group receiving DENA alone, a ZnONPs (10 μg/kg per week, intravenous (i.v.) for 1 month) control group, and a ZnONPs treatment group (receiving ZnONPs + DENA). ZnONPs significantly reduced the elevated serum levels of HCC-related tumor markers alphafetoprotein and alpha-l-fucosidase and the apoptotic marker caspase-3 compared with the untreated HCC rats. In addition, treatment with ZnONPs significantly decreased the elevated levels of hepatocyte integrity and oxidative stress markers as compared with the untreated HCC control group. Furthermore, the histopathological study revealed anaplasia and fibrous degenerations which were significantly corrected by ZnONPs treatment. In conclusion, administration of ZnONPs exhibited a promising preclinical anticancer efficacy in HCC and could be considered as a novel strategy for the treatment HCC in clinical practices. © 2016 John Wiley & Sons Australia, Ltd.

  3. Effect of mono-(2-ethylhexyl) phthalate on human and mouse fetal testis: In vitro and in vivo approaches

    Energy Technology Data Exchange (ETDEWEB)

    Muczynski, V. [Univ. Paris Diderot, Sorbonne Paris Cité, Laboratory of Development of the Gonads, Unit of Stem Cells and Radiation, BP 6, 92265 Fontenay-aux-Roses (France); CEA, DSV, iRCM, SCSR, LDRG, 92265 Fontenay-aux-Roses (France); INSERM, Unité 967, F-92265, Fontenay aux Roses (France); Cravedi, J.P. [INRA, INP, Université de Toulouse, UMR1331 TOXALIM, F-31027, Toulouse (France); Lehraiki, A.; Levacher, C.; Moison, D.; Lecureuil, C.; Messiaen, S. [Univ. Paris Diderot, Sorbonne Paris Cité, Laboratory of Development of the Gonads, Unit of Stem Cells and Radiation, BP 6, 92265 Fontenay-aux-Roses (France); CEA, DSV, iRCM, SCSR, LDRG, 92265 Fontenay-aux-Roses (France); INSERM, Unité 967, F-92265, Fontenay aux Roses (France); Perdu, E. [INRA, INP, Université de Toulouse, UMR1331 TOXALIM, F-31027, Toulouse (France); Frydman, R. [Service de Gynécologie-Obstétrique, Hôpital A. Béclère, Université Paris Sud F-92141 Clamart (France); Habert, R. [Univ. Paris Diderot, Sorbonne Paris Cité, Laboratory of Development of the Gonads, Unit of Stem Cells and Radiation, BP 6, 92265 Fontenay-aux-Roses (France); CEA, DSV, iRCM, SCSR, LDRG, 92265 Fontenay-aux-Roses (France); INSERM, Unité 967, F-92265, Fontenay aux Roses (France); and others

    2012-05-15

    The present study was conducted to determine whether exposure to the mono-(2-ethylhexyl) phthalate (MEHP) represents a genuine threat to male human reproductive function. To this aim, we investigated the effects on human male fetal germ cells of a 10{sup −5} M exposure. This dose is slightly above the mean concentrations found in human fetal cord blood samples by biomonitoring studies. The in vitro experimental approach was further validated for phthalate toxicity assessment by comparing the effects of in vitro and in vivo exposure in mouse testes. Human fetal testes were recovered during the first trimester (7–12 weeks) of gestation and cultured in the presence or not of 10{sup −5} M MEHP for three days. Apoptosis was quantified by measuring the percentage of Caspase-3 positive germ cells. The concentration of phthalate reaching the fetal gonads was determined by radioactivity measurements, after incubations with {sup 14}C-MEHP. A 10{sup −5} M exposure significantly increased the rate of apoptosis in human male fetal germ cells. The intratesticular MEHP concentration measured corresponded to the concentration added in vitro to the culture medium. Furthermore, a comparable effect on germ cell apoptosis in mouse fetal testes was induced both in vitro and in vivo. This study suggests that this 10{sup −5} M exposure is sufficient to induce changes to the in vivo development of the human fetal male germ cells. -- Highlights: ► 10{sup −5} M of MEHP impairs germ cell development in the human fetal testis. ► Organotypic culture is a suitable approach to investigate phthalate effects in human. ► MEHP is not metabolized in the human fetal testis. ► In mice, MEHP triggers similar effects both in vivo and in vitro.

  4. Trypanothione reductase: a target protein for a combined in vitro and in silico screening approach.

    Science.gov (United States)

    Beig, Mathias; Oellien, Frank; Garoff, Linnéa; Noack, Sandra; Krauth-Siegel, R Luise; Selzer, Paul M

    2015-01-01

    With the goal to identify novel trypanothione reductase (TR) inhibitors, we performed a combination of in vitro and in silico screening approaches. Starting from a highly diverse compound set of 2,816 compounds, 21 novel TR inhibiting compounds could be identified in the initial in vitro screening campaign against T. cruzi TR. All 21 in vitro hits were used in a subsequent similarity search-based in silico screening on a database containing 200,000 physically available compounds. The similarity search resulted in a data set containing 1,204 potential TR inhibitors, which was subjected to a second in vitro screening campaign leading to 61 additional active compounds. This corresponds to an approximately 10-fold enrichment compared to the initial pure in vitro screening. In total, 82 novel TR inhibitors with activities down to the nM range could be identified proving the validity of our combined in vitro/in silico approach. Moreover, the four most active compounds, showing IC50 values of <1 μM, were selected for determining the inhibitor constant. In first on parasites assays, three compounds inhibited the proliferation of bloodstream T. brucei cell line 449 with EC50 values down to 2 μM.

  5. Investigation of the in vitro culture process for skeletal-tissue-engineered constructs using computational fluid dynamics and experimental methods.

    Science.gov (United States)

    Hossain, Md Shakhawath; Chen, X B; Bergstrom, D J

    2012-12-01

    The in vitro culture process via bioreactors is critical to create tissue-engineered constructs (TECs) to repair or replace the damaged tissues/organs in various engineered applications. In the past, the TEC culture process was typically treated as a black box and performed on the basis of trial and error. Recently, computational fluid dynamics (CFD) has demonstrated its potential to analyze the fluid flow inside and around the TECs, therefore, being able to provide insight into the culture process, such as information on the velocity field and shear stress distribution that can significantly affect such cellular activities as cell viability and proliferation during the culture process. This paper briefly reviews the CFD and experimental methods used to investigate the in vitro culture process of skeletal-type TECs in bioreactors, where mechanical deformation of the TEC can be ignored. Specifically, this paper presents CFD modeling approaches for the analysis of the velocity and shear stress fields, mass transfer, and cell growth during the culture process and also describes various particle image velocimetry (PIV) based experimental methods to measure the velocity and shear stress in the in vitro culture process. Some key issues and challenges are also identified and discussed along with recommendations for future research.

  6. In vitro and in silico Approaches to Study Cytochrome P450-Mediated Interactions.

    Science.gov (United States)

    Tan, Boon Hooi; Pan, Yan; Dong, Amelia Nathania; Ong, Chin Eng

    2017-01-01

    In vitro and in silico models of drug metabolism are utilized regularly in the drug research and development as tools for assessing pharmacokinetic variability and drug-drug interaction risk. The use of in vitro and in silico predictive approaches offers advantages including guiding rational design of clinical drug-drug interaction studies, minimization of human risk in the clinical trials, as well as cost and time savings due to lesser attrition during compound development process. This article gives a review of some of the current in vitro and in silico methods used to characterize cytochrome P450(CYP)-mediated drug metabolism for estimating pharmacokinetic variability and the magnitude of drug-drug interactions. Examples demonstrating the predictive applicability of specific in vitro and in silico approaches are described. Commonly encountered confounding factors and sources of bias and error in these approaches are presented. With the advent of technological advancement in high throughput screening and computer power, the in vitro and in silico methods are becoming more efficient and reliable and will continue to contribute to the process of drug discovery, development and ultimately safer and more effective pharmacotherapy. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.

  7. A trivalent approach for determining in vitro toxicology: Examination of oxime K027.

    Science.gov (United States)

    Prado, Adriana; Petroianu, Georg A; Lorke, Dietrich E; Chambers, Jeremy W

    2015-02-01

    Unforeseen toxic effects contribute to compound attrition during preclinical evaluation and clinical trials. Consequently, there is a need to correlate in vitro toxicity to in vivo and clinical outcomes quickly and effectively. We propose an expedited evaluation of physiological parameters in vitro that will improve the ability to predict in vivo toxicity of potential therapeutics. By monitoring metabolism, mitochondrial physiology and cell viability, our approach provides insight to the extent of drug toxicity in vitro. To implement our approach, we used human hepatocellular carcinoma cells (HepG2) and neuroblastoma cells (SH-SY5Y) to monitor hepato- and neurotoxicity of the experimental oxime K027. We utilized a trivalent approach to measure metabolism, mitochondrial stress and induction of apoptosis in 96-well formats. Any change in these three areas may suggest drug-induced toxicity in vivo. K027 and pralidoxime, an oxime currently in clinical use, had no effect on glycolysis or oxygen consumption in HepG2 and SH-SY5Y cells. Similarly, these oximes did not induce oxidant generation nor alter mitochondrial membrane potential. Further, K027 and pralidoxime failed to activate effector caspases, and these oximes did not alter viability. The chemotherapeutic agent, docetaxel, negatively affected metabolism, mitochondrial physiology and viability. Our studies present a streamlined high-throughput trivalent approach for predicting toxicity in vitro, and this approach reveals that K027 has no measurable hepatotoxicity or neurotoxicity in vitro, which correlates with their in vivo data. This approach could eliminate toxic drugs from consideration for in vivo preclinical evaluation faster than existing toxicity prediction panels and ultimately prevent unnecessary experimentation. Copyright © 2014 John Wiley & Sons, Ltd.

  8. Antimicrobial activity of Tachyplesin 1 against Burkholderia pseudomallei: an in vitro and in silico approach

    Directory of Open Access Journals (Sweden)

    Lyn-Fay Lee

    2016-10-01

    Full Text Available Burkholderia pseudomallei, the causative agent of melioidosis, is intrinsically resistant to many conventional antibiotics. Therefore, alternative antimicrobial agents such as antimicrobial peptides (AMPs are extensively studied to combat this issue. Our study aims to identify and understand the mode of action of the potential AMP(s that are effective against B. pseudomallei in both planktonic and biofilm state as well as to predict the possible binding targets on using in vitro and in silico approaches. In the in vitro study, 11 AMPs were tested against 100 B. pseudomallei isolates for planktonic cell susceptibility, where LL-37, and PG1, demonstrated 100.0% susceptibility and TP1 demonstrated 83% susceptibility. Since the B. pseudomallei activity was reported on LL-37 and PG1, TP1 was selected for further investigation. TP1 inhibited B. pseudomallei cells at 61.69 μM, and membrane blebbing was observed using scanning electron microscopy. Moreover, TP1 inhibited B. pseudomallei cell growth, reaching bactericidal endpoint within 2 h post exposure as compared to ceftazidime (CAZ (8 h. Furthermore, TP1 was shown to suppress the growth of B. pseudomallei cells in biofilm state at concentrations above 221 μM. However, TP1 was cytotoxic to the mammalian cell lines tested. In the in silico study, molecular docking revealed that TP1 demonstrated a strong interaction to the common peptide or inhibitor binding targets for lipopolysaccharide of Escherichia coli, as well as autolysin, pneumolysin, and pneumococcal surface protein A (PspA of Streptococcus pneumoniae. Homology modelled B. pseudomallei PspA protein (YDP also showed a favourable binding with a strong electrostatic contribution and nine hydrogen bonds. In conclusion, TP1 demonstrated a good potential as an anti-B. pseudomallei agent.

  9. Investigation of tumor hypoxia using a two-enzyme system for in vitro generation of oxygen deficiency

    Directory of Open Access Journals (Sweden)

    Rana Shoaib

    2011-04-01

    Full Text Available Abstract Background Oxygen deficiency in tumor tissue is associated with a malign phenotype, characterized by high invasiveness, increased metastatic potential and poor prognosis. Hypoxia chambers are the established standard model for in vitro studies on tumor hypoxia. An enzymatic hypoxia system (GOX/CAT based on the use of glucose oxidase (GOX and catalase (CAT that allows induction of stable hypoxia for in vitro approaches more rapidly and with less operating expense has been introduced recently. Aim of this work is to compare the enzymatic system with the established technique of hypoxia chamber in respect of gene expression, glucose metabolism and radioresistance, prior to its application for in vitro investigation of oxygen deficiency. Methods Human head and neck squamous cell carcinoma HNO97 cells were incubated under normoxic and hypoxic conditions using both hypoxia chamber and the enzymatic model. Gene expression was investigated using Agilent microarray chips and real time PCR analysis. 14C-fluoro-deoxy-glucose uptake experiments were performed in order to evaluate cellular metabolism. Cell proliferation after photon irradiation was investigated for evaluation of radioresistance under normoxia and hypoxia using both a hypoxia chamber and the enzymatic system. Results The microarray analysis revealed a similar trend in the expression of known HIF-1 target genes between the two hypoxia systems for HNO97 cells. Quantitative RT-PCR demonstrated different kinetic patterns in the expression of carbonic anhydrase IX and lysyl oxidase, which might be due to the faster induction of hypoxia by the enzymatic system. 14C-fluoro-deoxy-glucose uptake assays showed a higher glucose metabolism under hypoxic conditions, especially for the enzymatic system. Proliferation experiments after photon irradiation revealed increased survival rates for the enzymatic model compared to hypoxia chamber and normoxia, indicating enhanced resistance to irradiation

  10. In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus: a biotechnological approach to study pine wilt disease.

    Science.gov (United States)

    Faria, Jorge M S; Sena, Inês; Vieira da Silva, Inês; Ribeiro, Bruno; Barbosa, Pedro; Ascensão, Lia; Bennett, Richard N; Mota, Manuel; Figueiredo, A Cristina

    2015-06-01

    Co-cultures of Pinus pinaster with Bursaphelenchus xylophilus were established as a biotechnological tool to evaluate the effect of nematotoxics addition in a host/parasite culture system. The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected for the first time in Europe in 1999 spreading throughout the pine forests in Portugal and recently in Spain. Plant in vitro cultures may be a useful experimental system to investigate the plant/nematode relationships in loco, thus avoiding the difficulties of field assays. In this study, Pinus pinaster in vitro cultures were established and compared to in vivo 1 year-old plantlets by analyzing shoot structure and volatiles production. In vitro co-cultures were established with the PWN and the effect of the phytoparasite on in vitro shoot structure, water content and volatiles production was evaluated. In vitro shoots showed similar structure and volatiles production to in vivo maritime pine plantlets. The first macroscopic symptoms of PWD were observed about 4 weeks after in vitro co-culture establishment. Nematode population in the culture medium increased and PWNs were detected in gaps of the callus tissue and in cavities developed from the degradation of cambial cells. In terms of volatiles main components, plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus co-cultures were all β- and α-pinene rich. Co-cultures may be an easy-to-handle biotechnological approach to study this pathology, envisioning the understanding of and finding ways to restrain this highly devastating nematode.

  11. In Vitro Investigation of D- and Lenantiomer Synergistic Efects of Some Amino Acids

    Directory of Open Access Journals (Sweden)

    Bystrická Zuzana

    2016-06-01

    Full Text Available D-amino acids can arise from endogenous microbial flora, from ingestion with the diet or from spontaneous racemization of L-amino acids during ageing. In this work, the behavior of methionine, homocysteine and cysteine enantiomers was investigated in human serum in vitro during 0-72 h at incubation temperature 37°C. The separation of enantiomers was realized in two dimensional on-line system (the connection of an achiral column Purospher RP-18 endcapped and a chiral column Chirobiotic TAG. This system allowed simultaneous monitoring all tested amino acids and their enantiomers. The possible effect D-enantiomer on the behavior of its L-enantiomer (the synergistic effect was evaluated during incubation time. The first results have showed that no synergistic effect of D-enantiomer on its Lisomer has been observed in our experimental conditions in vitro.

  12. Development of novel tools for the in vitro investigation of drug-induced liver injury.

    Science.gov (United States)

    Jiang, Jian; Wolters, Jarno E J; van Breda, Simone G; Kleinjans, Jos C; de Kok, Theo M

    2015-01-01

    Due to its complex mechanisms and unpredictable occurrence, drug-induced liver injury (DILI) complicates drug identification and classification. Since species-specific differences in metabolism and pharmacokinetics exist, data obtained from animal studies may not be sufficient to predict DILI in humans. Over the last few decades, numerous in vitro models have been developed to replace animal testing. The advantages and disadvantages of commonly used liver-derived in vitro models (e.g., cell lines, hepatocyte models, liver slices, three-dimensional (3D) hepatospheres, etc.) are discussed. Toxicogenomics-based methodologies (genomics, epigenomics, transcriptomics, proteomics and metabolomics) and next-generation sequencing have also been used to enhance the reliability of DILI prediction. This review presents an overview of the currently used alternative toxicological models and of the most advanced approaches in the field of DILI research. It seems unlikely that a single in vitro system will be able to mimic the complex interactions in the human liver. Three-dimensional multicellular systems may bridge the gap between conventional 2D models and in vivo clinical studies in humans and provide a reliable basis for hepatic toxicity assay development. Next-generation sequencing technologies, in comparison to microarray-based technologies, may overcome the current limitations and are promising for the development of predictive models in the near future.

  13. Characterizing Rat PNS Electrophysiological Response to Electrical Stimulation Using in vitro Chip-Based Human Investigational Platform (iCHIP)

    Energy Technology Data Exchange (ETDEWEB)

    Khani, Joshua [Georgetown Univ., Washington, DC (United States); Prescod, Lindsay [Georgetown Univ., Washington, DC (United States); Enright, Heather [Georgetown Univ., Washington, DC (United States); Felix, Sarah [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Osburn, Joanne [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Wheeler, Elizabeth [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Kulp, Kris [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-08-18

    Ex vivo systems and organ-on-a-chip technology offer an unprecedented approach to modeling the inner workings of the human body. The ultimate goal of LLNL’s in vitro Chip-based Human Investigational Platform (iCHIP) is to integrate multiple organ tissue cultures using microfluidic channels, multi-electrode arrays (MEA), and other biosensors in order to effectively simulate and study the responses and interactions of the major organs to chemical and physical stimulation. In this study, we focused on the peripheral nervous system (PNS) component of the iCHIP system. Specifically we sought to expound on prior research investigating the electrophysiological response of rat dorsal root ganglion cells (rDRGs) to chemical exposures, such as capsaicin. Our aim was to establish a protocol for electrical stimulation using the iCHIP device that would reliably elicit a characteristic response in rDRGs. By varying the parameters for both the stimulation properties – amplitude, phase width, phase shape, and stimulation/ return configuration – and the culture conditions – day in vitro and neural cell types - we were able to make several key observations and uncover a potential convention with a minimal number of devices tested. Future work will seek to establish a standard protocol for human DRGs in the iCHIP which will afford a portable, rapid method for determining the effects of toxins and novel therapeutics on the PNS.

  14. The Increase of Critical Thinking Skills through Mathematical Investigation Approach

    Science.gov (United States)

    Sumarna, N.; Wahyudin; Herman, T.

    2017-02-01

    Some research findings on critical thinking skills of prospective elementary teachers, showed a response that is not optimal. On the other hand, critical thinking skills will lead a student in the process of analysis, evaluation and synthesis in solving a mathematical problem. This study attempts to perform an alternative solution with a focus on mathematics learning conditions that is held in the lecture room through mathematical investigation approach. This research method was Quasi-Experimental design with pre-test post-test design. Data analysis using a mixed method with Embedded design. Subjects were regular students enrolled in 2014 at the study program of education of primary school teachers. The number of research subjects were 111 students consisting of 56 students in the experimental group and 55 students in the control group. The results of the study showed that (1) there is a significant difference in the improvement of critical thinking ability of students who receive learning through mathematical investigation approach when compared with students studying through expository approach, and (2) there is no interaction effect between prior knowledge of mathematics and learning factors (mathematical investigation and expository) to increase of critical thinking skills of students.

  15. Echinoderm regeneration: an in vitro approach using the crinoid Antedon mediterranea.

    Science.gov (United States)

    Di Benedetto, Cristiano; Parma, Lorenzo; Barbaglio, Alice; Sugni, Michela; Bonasoro, Francesco; Carnevali, Maria Daniela Candia

    2014-10-01

    Among echinoderms, crinoids are well known for their remarkable regenerative potential. Regeneration depends mainly on progenitor cells (undifferentiated or differentiated), which migrate and proliferate in the lesion site. The crucial role of the "progenitor" elements involved in the regenerative processes, in terms of cell recruitment, sources, and fate, is a central problem in view of its topical interest and biological implications. The spectacular regenerative potential of crinoids is used to replace lost internal and external organs. In particular, the process of arm regeneration in the feather star Antedon mediterranea is the regeneration model most extensively explored to date. We have addressed the morphological and functional characterization of the cell phenotypes responsible for the arm regenerative processes by using an in vitro approach. This represents the first successful attempt to culture cells involved in crinoid regeneration. A comparison of these results with others from previous in vivo investigations confirms the diverse cell types contributing to regeneration and underscores their involvement in migration, proliferation, and dedifferentiation processes.

  16. Curricular approaches to research ethics training for psychiatric investigators.

    Science.gov (United States)

    Chen, Donna T

    2003-12-01

    Training in research ethics is crucial for psychiatric investigators. Addressing ethical dimensions of human subjects research requires knowledge about the rules and norms governing research; sensitivity to ethical implications of actions; and skills in ethics problem solving. Investigators in training who are physicians have the additional challenge of developing identities as investigators that sometimes conflict with their identities as physicians. To propose a curriculum for psychiatric research ethics training. Review of literature on ethics education and presentation of a curricular approach to research ethics training for psychiatric investigators. Research ethics can be learned and should be taught. Involvement of active investigators in teaching research ethics is important. While core topics of psychiatric research ethics training have not yet been identified, there are available models from which to draw. Research ethics should be introduced early and integrated throughout the research training period. Lack of resources and institutional support can be obstacles to development of comprehensive research ethics curricula. Small-group, case-based discussion is best for teaching ethics problem-solving skills. Examples of teaching module ideas and a fully developed sample teaching module are presented. There is opportunity for creative models for teaching psychiatric research ethics. Work is needed to identify core topics, target pedagogical strategies to trainees at different levels, and develop evaluation methods.

  17. High-throughput, cell-free, liposome-based approach for assessing in vitro activity of lipid kinases.

    Science.gov (United States)

    Demian, Douglas J; Clugston, Susan L; Foster, Meta M; Rameh, Lucia; Sarkes, Deborah; Townson, Sharon A; Yang, Lily; Zhang, Melvin; Charlton, Maura E

    2009-08-01

    Lipid kinases are central players in lipid signaling pathways involved in inflammation, tumorigenesis, and metabolic syndrome. A number of these kinase targets have proven difficult to investigate in higher throughput cell-free assay systems. This challenge is partially due to specific substrate interaction requirements for several of the lipid kinase family members and the resulting incompatibility of these substrates with most established, homogeneous assay formats. Traditional, cell-free in vitro investigational methods for members of the lipid kinase family typically involve substrate incorporation of [gamma-32P] and resolution of signal by thin-layer chromatography (TLC) and autoradiograph densitometry. This approach, although highly sensitive, does not lend itself to high-throughput testing of large numbers of small molecules (100 s to 1 MM+). The authors present the development and implementation of a fully synthetic, liposome-based assay for assessing in vitro activity of phosphatidylinositol-5-phosphate-4-kinase isoforms (PIP4KIIbeta and alpha) in 2 commonly used homogeneous technologies. They have validated these assays through compound testing in both traditional TLC and radioactive filterplate approaches as well as binding validation using isothermic calorimetry. A directed library representing known kinase pharmacophores was screened against type IIbeta phosphatidylinositol-phosphate kinase (PIPK) to identify small-molecule inhibitors. This assay system can be applied to other types and isoforms of PIPKs as well as a variety of other lipid kinase targets.

  18. Clozapine-induced agranulocytosis: Evidence for an immune-mediated mechanism from a patient-specific in-vitro approach.

    Science.gov (United States)

    Regen, Francesca; Herzog, Irmelin; Hahn, Eric; Ruehl, Claudia; Le Bret, Nathalie; Dettling, Michael; Heuser, Isabella; Hellmann-Regen, Julian

    2017-02-01

    Use of the atypical antipsychotic clozapine (CZP) is compromised by the risk of potentially fatal agranulocytosis/granulocytopenia (CIAG). To address this, we have established a simple, personalized cell culture-based strategy to identify CIAG-susceptible patients, hypothesizing that an immunogenic and possibly haptene-based mechanism underlies CIAG pathophysiology. To detect a putative haptene-induced response to CZP in vitro exposure, a traditional lymphocyte stimulation assay was adapted and applied to patient-specific peripheral blood-derived mononuclear cells (PBMC). 6 patients with a history of CIAG, 6 patients under CZP treatment (without CIAG) and 12 matched healthy controls were studied. In vitro CZP exposure, even at strikingly low levels, resulted in significantly increased proliferation rates only in CIAG patients' PBMC. Other parameters including cell viability and mitogen-induced proliferation were also affected by in vitro CZP exposure, yet there was no significant difference between the groups. This personalized approach is a starting point for further investigations into a putative haptene-based mechanism underlying CIAG development, and may facilitate the future development of predictive testing. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Investigation of a novel approach for aquaculture site selection.

    Science.gov (United States)

    Falconer, Lynne; Telfer, Trevor C; Ross, Lindsay G

    2016-10-01

    This study investigated the potential use of two "species distribution models" (SDMs), Mahalanobis Typicality and Maxent, for aquaculture site selection. SDMs are used in ecological studies to predict the spatial distribution of species based on analysis of conditions at locations of known presence or absence. Here the input points are aquaculture sites, rather than species occurrence, thus the models evaluate the parameters at the sites and identify similar areas across the rest of the study area. This is a novel approach that avoids the need for data reclassification and weighting which can be a source of conflict and uncertainty within the commonly used multi-criteria evaluation (MCE) technique. Using pangasius culture in the Mekong Delta, Vietnam, as a case study, Mahalanobis Typicality and Maxent SDMs were evaluated against two models developed using the MCE approach. Mahalanobis Typicality and Maxent assess suitability based on similarity to existing farms, while the MCE approach assesses suitability using optimal values for culture. Mahalanobis Typicality considers the variables to have equal importance whereas Maxent analyses the variables to determine those which influence the distribution of the input data. All of the models indicate there are suitable areas for culture along the two main channels of the Mekong River which are currently used to farm pangasius and also inland in the north and east of the study area. The results show the Mahalanobis Typicality model had more high scoring areas and greater overall similarity than Maxent to the MCE outputs, suggesting, for this case study, it was the most appropriate SDM for aquaculture site selection. With suitable input data, a combined SDM and MCE model would overcome limitations of the individual approaches, allowing more robust planning and management decisions for aquaculture, other stakeholders and the environment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Combinational approach using solid dispersion and semi-solid matrix technology to enhance in vitro dissolution of telmisartan

    Directory of Open Access Journals (Sweden)

    Syed Faisal Ali

    2016-02-01

    Full Text Available The present investigation was focused to formulate semi-solid capsules (SSCs of hydrophobic drug telmisartan (TLMS by encapsulating semi-solid matrix of its solid dispersion (SD in HPMC capsules. The combinational approach was used to reduce the lag time in drug release and improvise its dissolution. SDs of TLMS was prepared using hot fusion method by varying the combinations of Pluronic-F68, Gelucire 50/13 and Plasdone S630. A total of nine batches (SD1-SD9 were characterized for micromeritic properties, in vitro dissolution behavior and surface characterization. SD4 with 52.43% cumulative drug release (CDR in phosphate buffer, pH 7.4, in 120 min, t50% 44.2 min and DE30min 96.76% was selected for the development of semi-solid capsules. Differential scanning calorimetry of SD4 revealed molecular dispersion of TLMS in Pluronic-F68. SD4 was formulated into SSCs using Gelucire 44/14 and PEG 400 as semi-solid components and PEG 6000 as a suspending agent to achieve reduction in lag time for effective drug dissolution. SSC6 showed maximum in vitro drug dissolution 97.49 % in phosphate buffer, pH 7.4 with in 20 min that was almost a three folds reduction in the time required to achieve similar dissolution by SD. Thus, SSCs present an excellent approach to enhance in vitro dissolution as well as to reduce the lag time of dissolution for poorly water soluble drugs especially to those therapeutic classes that are intended for faster onset of action. Developed approach based on HPMC capsules provided a better alternative to target delivery of telmisartan to the vegetarian population.

  1. Toxicological assessment of kretek cigarettes Part 4: mechanistic investigations, smoke chemistry and in vitro toxicity.

    Science.gov (United States)

    Roemer, E; Dempsey, R; Lawless-Pyne, J; Lukman, S; Evans, A Deger; Trelles-Sticken, E; Wittke, S; Schorp, M K

    2014-12-01

    The smoke chemistry and in vitro toxicity of mainstream smoke (MS) was investigated in American-blended cigarettes with or without the addition of 2.5%, 5% or 10% eugenol to the tobacco and in Indonesian-blended cigarettes with and without the addition of cloves, cloves extracted with hot ethanol, and extracted cloves replenished with eugenol or clove oil. The addition of eugenol reduced the concentration of nearly all toxicants measured in MS as well as the in vitro cytotoxicity of the gas/vapor phase. Reductions were also seen in bacterial mutagenicity of the total particulate matter (TPM) assessed by the Ames Assay. The addition of extracted cloves led to increases and decreases of toxicant concentrations in MS. Replenishment with eugenol or clove oil decreased the toxicant concentrations; with most smoke constituent concentrations reduced below the concentration found in tobacco-only cigarettes. Cytotoxicity of the TPM was not affected by the clove preparations. However, GVP cytotoxicity was reduced (untreated cloves showing the highest reductions). Mutagenicity of TPM was decreased by the clove preparations. Mechanisms for the reductions, (up to 40%), are most likely due to dilution effects by eugenol, changed burning characteristics of the tobacco, and free radical scavenging by eugenol. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Investigation of modification X-ray induced bystander effect in vitro.

    Science.gov (United States)

    Shemetun, O V; Talan, O O

    2014-09-01

    Objective - to investigate the modification of bystander effect induced by X-irradiation of human peripheral blood in vitro by application of antioxidant vitamin medication. Material and methods. Modeling of radiation-induced bystander effect in vitro in mixed lymphocyte cultures exposed to dose of 1 Gy and non-irradiated blood lymphocytes of persons of different sexes, GTG-staining of metaphase chromosomes and their cytogenetic analysis; application of antioxidant preparation (soluble forms of vitamins E, C and A) in concentration 40 μg/ml. Results. Under the introduction of antioxidant preparation into mixed culture before lymphocytes cultivation frequency of chromosomal aberrations in bystander cells did not significantly different from the control (p > 0.05). application of antioxidant preparation modifies the radiation-induced bystander effect in unirradiated human peripheral blood lymphocytes under their joint cultivation with lymphocytes irradiated in dose of 1 Gy. Antioxidant prevents the development of secondary oxidative stress in unirradiated cells, eliminates the development in them of radiation-induced bystander effect and ensures the preservation of stability of their chromosome apparatus. O. V. Shemetun, O. O. Talan.

  3. Investigating the ocular temperature rise during femtosecond laser lens fragmentation: an in vitro study.

    Science.gov (United States)

    Mencucci, Rita; Matteoli, Sara; Corvi, Andrea; Terracciano, Luca; Favuzza, Eleonora; Gherardini, Stefano; Caruso, Filippo; Bellucci, Roberto

    2015-12-01

    To investigate the trend of temperature variation during lens fragmentation simulated by a femtosecond laser on an in vitro eye model. In our experimental study, a convex cylinder of gelatinous material, usually employed in femtosecond laser calibration, was used to simulate both an anterior segment and a crystalline lens during fragmentation performed with the Victus femtosecond laser (Technolas Perfect Vision GmbH, Germany; Bausch + Lomb Incorporated, USA). Two radiated energies (7000 nJ and 9000 nJ) and three cutting patterns (crosses, circles and cross + circle) were applied. Trends of temperature variation as a function of time were obtained using a T-type thermocouple. The maximum value of temperature rise during lens fragmentation ranged from 3.53 to 5.13 °C; the rise was directly proportional to the intensity of the radiated energy (7000 nJ or 9000 nJ) and the cutting pattern performed. This behavior was experimentally represented by an asymmetric function with a characteristic bell curve shape, whereas it was mathematically described by a transport diffusive model. Since the temperature rise at the fragmentation volume base resulted to be around 5 °C in our in vitro study, lens fragmentation performed using the Victus femtosecond laser might be considered safe form a thermal point of view.

  4. In vitro bioequivalence approach for a locally acting gastrointestinal drug: lanthanum carbonate.

    Science.gov (United States)

    Yang, Yongsheng; Shah, Rakhi B; Yu, Lawrence X; Khan, Mansoor A

    2013-02-04

    A conventional human pharmacokinetic (PK) in vivo study is often considered as the "gold standard" to determine bioequivalence (BE) of drug products. However, this BE approach is not always applicable to the products not intended to be delivered into the systemic circulation. For locally acting gastrointestinal (GI) products, well designed in vitro approaches might be more practical in that they are able not only to qualitatively predict the presence of the active substance at the site of action but also to specifically assess the performance of the active substance. For example, lanthanum carbonate chewable tablet, a locally acting GI phosphate binder when orally administrated, can release free lanthanum ions in the acid environment of the upper GI tract. The lanthanum ions directly reach the site of action to bind with dietary phosphate released from food to form highly insoluble lanthanum-phosphate complexes. This prevents the absorption of phosphate consequently reducing the serum phosphate. Thus, using a conventional PK approach to demonstrate BE is meaningless since plasma levels are not relevant for local efficacy in the GI tract. Additionally the bioavailability of lanthanum carbonate is less than 0.002%, and therefore, the PK approach is not feasible. Therefore, an alternative assessment method is required. This paper presents an in vitro approach that can be used in lieu of PK or clinical studies to determine the BE of lanthanum carbonate chewable tablets. It is hoped that this information can be used to finalize an in vitro guidance for BE studies of lanthanum carbonate chewable tablets as well as to assist with "in vivo" biowaiver decision making. The scientific information might be useful to the pharmaceutical industry for the purpose of planning and designing future BE studies.

  5. In vitro manipulation of gene expression in larval Schistosoma: a model for postgenomic approaches in Trematoda

    Science.gov (United States)

    YOSHINO, TIMOTHY P.; DINGUIRARD, NATHALIE; DE MORAES MOURÃO, MARINA

    2013-01-01

    SUMMARY With rapid developments in DNA and protein sequencing technologies, combined with powerful bioinformatics tools, a continued acceleration of gene identification in parasitic helminths is predicted, potentially leading to discovery of new drug and vaccine targets, enhanced diagnostics and insights into the complex biology underlying host-parasite interactions. For the schistosome blood flukes, with the recent completion of genome sequencing and comprehensive transcriptomic datasets, there has accumulated massive amounts of gene sequence data, for which, in the vast majority of cases, little is known about actual functions within the intact organism. In this review we attempt to bring together traditional in vitro cultivation approaches and recent emergent technologies of molecular genomics, transcriptomics and genetic manipulation to illustrate the considerable progress made in our understanding of trematode gene expression and function during development of the intramolluscan larval stages. Using several prominent trematode families (Schistosomatidae, Fasciolidae, Echinostomatidae), we have focused on the current status of in vitro larval isolation/cultivation as a source of valuable raw material supporting gene discovery efforts in model digeneans that include whole genome sequencing, transcript and protein expression profiling during larval development, and progress made in the in vitro manipulation of genes and their expression in larval trematodes using transgenic and RNA interference (RNAi) approaches. PMID:19961646

  6. An in vitro approach for comparative interspecies metabolism of agrochemicals.

    Science.gov (United States)

    Whalley, Paul M; Bartels, Michael; Bentley, Karin S; Corvaro, Marco; Funk, Dorothee; Himmelstein, Matthew W; Neumann, Birgit; Strupp, Christian; Zhang, Fagen; Mehta, Jyotigna

    2017-08-01

    The metabolism and elimination of a xenobiotic has a direct bearing on its potential to cause toxicity in an organism. The confidence with which data from safety studies can be extrapolated to humans depends, among other factors, upon knowing whether humans are systemically exposed to the same chemical entities (i.e. a parent compound and its metabolites) as the laboratory animals used to study toxicity. Ideally, to understand a metabolite in terms of safety, both the chemical structure and the systemic exposure would need to be determined. However, as systemic exposure data (i.e. blood concentration/time data of test material or metabolites) in humans will not be available for agrochemicals, an in vitro approach must be taken. This paper outlines an in vitro experimental approach for evaluating interspecies metabolic comparisons between humans and animal species used in safety studies. The aim is to ensure, where possible, that all potential human metabolites are also present in the species used in the safety studies. If a metabolite is only observed in human in vitro samples and is not present in a metabolic pathway defined in the toxicological species already, the toxicological relevance of this metabolite must be evaluated. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  7. Evaluating in Vitro Culture Medium of Gut Microbiome with Orthogonal Experimental Design and a Metaproteomics Approach.

    Science.gov (United States)

    Li, Leyuan; Zhang, Xu; Ning, Zhibin; Mayne, Janice; Moore, Jasmine I; Butcher, James; Chiang, Cheng-Kang; Mack, David; Stintzi, Alain; Figeys, Daniel

    2017-11-22

    In vitro culture based approaches are time- and cost-effective solutions for rapidly evaluating the effects of drugs or natural compounds against microbiomes. The nutritional composition of the culture medium is an important determinant for effectively maintaining the gut microbiome in vitro. This study combines orthogonal experimental design and a metaproteomics approach to obtaining functional insights into the effects of different medium components on the microbiome. Our results show that the metaproteomic profile respond differently to medium components, including inorganic salts, bile salts, mucin, and short-chain fatty acids. Multifactor analysis of variance further revealed significant main and interaction effects of inorganic salts, bile salts, and mucin on the different functional groups of gut microbial proteins. While a broad regulating effect was observed on basic metabolic pathways, different medium components also showed significant modulations on cell wall, membrane, and envelope biogenesis and cell motility related functions. In particular, flagellar assembly related proteins were significantly responsive to the presence of mucin. This study provides information on the functional influences of medium components on the in vitro growth of microbiome communities and gives insight on the key components that must be considered when selecting and optimizing media for culturing ex vivo microbiotas.

  8. Biodegradable polymeric system for cisplatin delivery: Development, in vitro characterization and investigation of toxicity profile

    Energy Technology Data Exchange (ETDEWEB)

    Alam, Noor; Khare, Vaibhav; Dubey, Ravindra; Saneja, Ankit [Formulation and Drug Delivery Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Kushwaha, Manoj; Singh, Gurdarshan; Sharma, Neelam; Chandan, Balkrishan [PK-PD-Toxicology Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India); Gupta, Prem N., E-mail: pngupta10@gmail.com [Formulation and Drug Delivery Division, CSIR-Indian Institute of Integrative Medicine, Jammu (India)

    2014-05-01

    Cisplatin is one of the most potent anticancer agent used in the treatment of various solid tumors, however, its clinical use is limited due to severe adverse effects including nephrotoxicity. In this investigation cisplatin loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles were developed and characterized for various in vitro characteristics including size distribution, zeta potential, drug loading and release profile. PLGA nanoparticles were successfully developed as investigated using scanning electron microscopy and exhibited average particles size and zeta potential as 284.8 nm and − 15.8 mV, respectively. Fourier transform infrared spectroscopy and differential scanning calorimetry indicated an absence of any polymer–drug interactions. Cisplatin nanoparticles exhibited in vitro anticancer activity against A549 cells comparable to that of cisplatin solution. The biodistribution study in mice indicated that the kidney cisplatin level was significantly (p < 0.01) lower with cisplatin nanoparticles than cisplatin solution. Following two cycles of cisplatin treatment, a week apart, blood urea nitrogen level was found to be higher in case of cisplatin solution as compared to cisplatin nanoparticles. Further, there was a significant (p < 0.01) increase in plasma creatinine level in case of cisplatin solution as compared to cisplatin nanoparticles. Histopathological examination of kidney from cisplatin nanoparticles treated group revealed no kidney damage, however, a sign of nephrotoxicity was observed in the case of cisplatin solution. The results suggest that PLGA nanoparticle based formulation could be a potential option for cisplatin delivery. - Highlights: • Cisplatin is detected by LCMS following complexation with DDTC. • Nanoparticles showed lower cisplatin accumulation in the kidney. • Nephrotoxicity was evaluated by BUN and creatinine level and by histopathology. • Nanoparticles exhibited lower nephrotoxicity.

  9. Investigating the Research Approaches for Examining Technology Adoption Issues

    Directory of Open Access Journals (Sweden)

    Jyoti Choudrie

    2005-01-01

    Full Text Available Adoption of technology, a research topic within the Information Systems area, is usually studied at two levels: organizational level and user level. This paper examines the range of methods used for studying technology adoption issues at both these levels. The approaches were selected after conducting a review of 48 articles on technology adoption and usage, published in peer reviewed journals between 1985 and 2003. The journals reviewed include the MIS Quarterly, Information Systems Research, European Journal of Information Systems, Information Systems Journal, and other relevant journals in the IS area. The findings suggest that the survey method was used predominantly when investigating the topics of user adoption and the usage of technology. In contrast, the case study method is the most widely used when examining adoption issues at the organizational level.

  10. Investigation of an angular spectrum approach for pulsed ultrasound fields

    DEFF Research Database (Denmark)

    Du, Yigang; Jensen, Henrik; Jensen, Jørgen Arendt

    2013-01-01

    An Angular Spectrum Approach (ASA)is formulated and employed to simulate linear pulsed ultra sound fields for high bandwidth signals. Ageometrically focused piston transducer is used as the acoustic source. Signals are cross-correlated to findthe true sound speed during the measurement to make...... the simulated and measured pulses in phase for comparisons. The calculated sound speed in the measurement is varied between 1487.45 m/s and 1487.75 m/s by using different initial values in the ASA simulation. Results from the pulsed ASA simulation susing both Field II simulated and hydrophone measured acoustic...... sources are compared to the Field II simulated and hydroph one measure dpulses, respectively. The total relative root mean squar e(RMS)errors of the pulsed ASA are investigated by using different time-point, zero-padding factors, spatial sampling interval and temporal sampling frequency in the sim ulation...

  11. Approach to Investigating Congenital Skeletal Abnormalities in Livestock.

    Science.gov (United States)

    Dittmer, K E; Thompson, K G

    2015-09-01

    Congenital skeletal abnormalities may be genetic, teratogenic, or nutritional in origin; distinguishing among these different causes is essential in the management of the disease but may be challenging. In some cases, teratogenic or nutritional causes of skeletal abnormalities may appear very similar to genetic causes. For example, chondrodysplasia associated with intrauterine zinc or manganese deficiency and mild forms of hereditary chondrodysplasia have very similar clinical features and histologic lesions. Therefore, historical data are essential in any attempt to distinguish genetic and acquired causes of skeletal lesions; as many animals as possible should be examined; and samples should be collected for future analysis, such as genetic testing. Acquired causes of defects often show substantial variation in presentation and may improve with time, while genetic causes frequently have a consistent presentation. If a disease is determined to be of genetic origin, a number of approaches may be used to detect mutations, each with advantages and disadvantages. These approaches include sequencing candidate genes, single-nucleotide polymorphism array with genomewide association studies, and exome or whole genome sequencing. Despite advances in technology and increased cost-effectiveness of these techniques, a good clinical history and description of the pathology and a reliable diagnosis are still key components of any investigation. © The Author(s) 2015.

  12. Advantages of the basin approach for investigations of aufeises (naleds

    Directory of Open Access Journals (Sweden)

    V. V. Shepelev

    2016-01-01

    Full Text Available Formation of aufeises (naleds is rather widely distributed cryogenic process in the permafrost zone, therefore naleds are reasonably called the seasonal glaciation of the planet. The ever-increasing interest in aufeises significantly extends the thematic focus of studying them. Various methodological approaches (basin, geological-structural, lithologic-facies, statistical, etc. are used to reveal regularities of the aufeis occurrence. Using several river basins in Yakutia and its territory as the examples, the perspectivity of the basin approach for investigation of the aufeis distribution is substantiated in this study. The results demonstrated the clear relationship between the quantitative indicators of areal distribution of naleds and altitudes of places of their formation for individual river basins. This relationship suggests that aufeises in river basins of the permafrost zones are important elements in the water-balance and water exchange, which is closely interrelated with not only climatic and morphometric characteristics of a basin, but it does also correlate with its hydrological, hydrogeological, geological, geocryological, landscape and other conditions.

  13. Prion structure investigated in situ, ex vivo, and in vitro by FTIR spectroscopy

    Science.gov (United States)

    Kneipp, Janina; Miller, Lisa M.; Spassov, Sashko; Sokolowski, Fabian; Lasch, Peter; Beekes, Michael; Naumann, Dieter

    2004-07-01

    Syrian hamster nervous tissue was investigated by FTIR microspectroscopy with conventional and synchrotron infrared light sources. Various tissue structures from the cerebellum and medulla oblongata of scrapie-infected and control hamsters were investigated at a spatial resolution of 50 μm. Single neurons in dorsal root ganglia of scrapie-infected hamsters were analyzed by raster scan mapping at 6 μm spatial resolution. These measurements enabled us to (i) scrutinize structural differences between infected and non-infected tissue and (ii) analyze for the first time the distribution of different protein structures in situ within single nerve cells. Single nerve cells exhibited areas of increased β-sheet content, which co-localized consistently with accumulations of the pathological prion protein (PrPSc). Spectral data were also obtained from purified, partly proteinase K digested PrPSc isolated from scrapie-infected nervous tissue of hamsters to elucidate similarities/dissimilarities between prion structure in situ and ex vivo. A further comparison is drawn to the recombinant Syrian hamster prion protein SHaPrP90-232, whose in vitro transition from the predominantly a-helical isoform to β-sheet rich oligomeric structures was also investigated by FTIR spectroscopy.

  14. Summary of: an in vitro investigation of the erosive potential of smoothies.

    Science.gov (United States)

    Brand, H S

    2013-02-01

    Recent health promotion campaigns have encouraged the public to consume at least five portions of fruit and vegetables per day. Many see consuming fruit smoothies as a way of achieving this. To ascertain the potential or otherwise for fruit smoothies to bring about dental erosion. Laboratory study. This was an in vitro investigation in which five varieties of shop bought fruit smoothies, including a 'thickie' were investigated, with respect to their initial pH, titratable acidity and effect upon exposure to the surface microhardness and profile of extracted human teeth. In addition their performance was compared to negative (Volvic™ water) and positive (orange juice) control drinks as well as a homemade smoothie, based upon the recipe of one of the commercially bought drinks, from which ingredient omissions were made. The majority of the drinks investigated had a baseline pH below the critical pH of enamel (5.5) and required comparable volumes of 0.1M NaOH to raise their pH to neutrality as the positive control. Only two drinks (Volvic™ still mineral water, the negative control, and the yoghurt, vanilla bean and honey 'thickie') displayed a higher pH, though to neutralise the thickie, a lesser quantity of alkali addition was required. The immersion of the tooth samples in the drinks brought about reductions in their surface hardness (expressed as a percentage change of median hardness) but these were only significant (p benefits of their consumption, their consumption should be confined to mealtimes.

  15. Investigating causes of changes in runoff using hydrological simulation approach

    Science.gov (United States)

    Wang, Guoqing; Zhang, Jianyun; Li, Xuemei; Bao, Zhenxin; Liu, Yanli; Liu, Cuishan; He, Ruimin; Luo, Junsong

    2017-09-01

    Stream flow plays a crucial role in environmental, social and economic contexts. It is of significance to investigate the causes of change in runoff for better water resources management. This study detects the variation trend of recorded runoff of the Gushan River, a tributary of the Yellow River located on the Loess Plateau with severe soil and water losing, and investigates the impacts of climate change and human activities on runoff using hydrological simulation approach. Results show that the recorded runoff at Gaoshiya station on the Gushan River has experienced a significant declining trend from 1954-2013 with an abrupt change occurring in 1973. SimHyd rainfall runoff model performs well for monthly discharge simulation with Nash-Sutcliffe coefficient of 82.6 % and relative error of 0.32 %. Runoff depth over the catchment in 1980-2013 reduced by 52.4 mm compared to the previous period, in which human activities and climate change contribute 61.5 and 38.5 % of the total runoff reduction, respectively. However, the human-induced impact tends to increase. Therefore, efforts to improve the ecology of the Loess Plateau should give sufficient attention to the impacts of human activity.

  16. A cocktail approach for assessing the in vitro activity of human cytochrome P450s: an overview of current methodologies.

    Science.gov (United States)

    Spaggiari, Dany; Geiser, Laurent; Daali, Youssef; Rudaz, Serge

    2014-12-01

    An assessment of cytochrome P450 (CYP) enzyme activity is essential for characterizing the phase I metabolism of biological systems or to evaluate the inhibition/induction properties of xenobiotics. CYPs have generally been investigated individually by single probes, and metabolite formation has been monitored by liquid chromatography-mass spectrometry (LC-MS). To increase the throughput, many probes have been applied to assess multiple CYP activities simultaneously within a single experiment. This strategy is called the cocktail approach, and it has already been reviewed for in vivo applications, but never for in vitro ones. This review focuses for the first time on an in vitro cocktail approach, and it references the most notable articles on this topic. The advantages and limitations of applying cocktails for the in vitro activity assessment of major human CYPs, namely, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and subfamily CYP3A, are discussed. This article considers the probe reaction selections for each CYP according to regulatory recommendations, probe metabolic properties (i.e., specificity and turnover), probe concentrations and analytical sensitivity, but it also highlights a challenge specific to cocktail design, which is probe-probe interaction. The last part of the review reports some methodologies for incubating these cocktails and discusses some important issues regarding the incubation time, enzyme concentrations and sample preparation. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. [Investigation of in vitro metronidazole resistance in the clinical isolates of Trichomonas vaginalis].

    Science.gov (United States)

    Ertabaklar, Hatice; Yaman Karadam, Senem; Malatyalı, Erdoğan; Ertuğ, Sema

    2016-10-01

    -sensitive isolates were 27.17 µg/ml in aerobic and 7.75 µg/ml in anaerobic conditions. The rate of metronidazole resistance detected in this study was higher than most of reports from different countries. Despite being limited to the isolates from Aydin province (located at Agean region of Turkey), the present study has a value as it presented the existence of metronidazole-resistant isolates in Turkey for the first time. More research from other parts of Turkey is needed to better understand the metronidazole resistance at a national scale and to investigate novel strategies for the treatment. Moreover, further studies need to be carried out in order to clarify the relationship between clinical treatment response and in vitro metronidazole resistance in trichomoniasis.

  18. Patient-specific in vitro models for hemodynamic analysis of congenital heart disease - Additive manufacturing approach.

    Science.gov (United States)

    Medero, Rafael; García-Rodríguez, Sylvana; François, Christopher J; Roldán-Alzate, Alejandro

    2017-03-21

    Non-invasive hemodynamic assessment of total cavopulmonary connection (TCPC) is challenging due to the complex anatomy. Additive manufacturing (AM) is a suitable alternative for creating patient-specific in vitro models for flow measurements using four-dimensional (4D) Flow MRI. These in vitro systems have the potential to serve as validation for computational fluid dynamics (CFD), simulating different physiological conditions. This study investigated three different AM technologies, stereolithography (SLA), selective laser sintering (SLS) and fused deposition modeling (FDM), to determine differences in hemodynamics when measuring flow using 4D Flow MRI. The models were created using patient-specific MRI data from an extracardiac TCPC. These models were connected to a perfusion pump circulating water at three different flow rates. Data was processed for visualization and quantification of velocity, flow distribution, vorticity and kinetic energy. These results were compared between each model. In addition, the flow distribution obtained in vitro was compared to in vivo. The results showed significant difference in velocities measured at the outlets of the models that required internal support material when printing. Furthermore, an ultrasound flow sensor was used to validate flow measurements at the inlets and outlets of the in vitro models. These results were highly correlated to those measured with 4D Flow MRI. This study showed that commercially available AM technologies can be used to create patient-specific vascular models for in vitro hemodynamic studies at reasonable costs. However, technologies that do not require internal supports during manufacturing allow smoother internal surfaces, which makes them better suited for flow analyses. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Investigation of volatile organic biomarkers derived from Plasmodium falciparum in vitro

    Directory of Open Access Journals (Sweden)

    Wong Rina PM

    2012-09-01

    Full Text Available Abstract Background There remains a need for techniques that improve the sensitive detection of viable Plasmodium falciparum as part of diagnosis and therapeutic monitoring in clinical studies and usual-care management of malaria infections. A non-invasive breath test based on P. falciparum-associated specific volatile organic compounds (VOCs could fill this gap and provide insights into parasite metabolism and pathogenicity. The aim of this study was to determine whether VOCs are present in the headspace above in vitro P. falciparum cultures. Methods A novel, custom-designed apparatus was developed to enable efficient headspace sampling of infected and non-infected cultures. Conditions were optimized to support cultures of high parasitaemia (>20% to improve the potential detection of parasite-specific VOCs. A number of techniques for VOC analysis were investigated including solid phase micro-extraction using two different polarity fibres, and purge and trap/thermal desorption, each coupled to gas chromatography–mass spectrometry. Each experiment and analysis method was performed at least on two occasions. VOCs were identified by comparing their mass spectra against commercial mass spectral libraries. Results No unique malarial-specific VOCs could be detected relative to those in the control red blood cell cultures. This could reflect sequestration of VOCs into cell membranes and/or culture media but solvent extractions of supernatants and cell lysates using hexane, dichloromethane and ethyl acetate also showed no obvious difference compared to control non-parasitized cultures. Conclusions Future in vivo studies analysing the breath of patients with severe malaria who are harbouring a parasite biomass that is significantly greater than achievable in vitro may yet reveal specific clinically-useful volatile chemical biomarkers.

  20. In vitro performance investigation of bioresorbable scaffolds – Standard tests for vascular stents and beyond

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, Wolfram, E-mail: wolfram.schmidt@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Behrens, Peter, E-mail: peter.behrens@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Brandt-Wunderlich, Christoph, E-mail: christoph.brandt@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Siewert, Stefan, E-mail: stefan.siewert@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Grabow, Niels, E-mail: niels.grabow@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Schmitz, Klaus-Peter, E-mail: klaus-peter.schmitz@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany)

    2016-09-15

    Background/Purpose: Biodegradable polymers are the main materials for coronary scaffolds. Magnesium has been investigated as a potential alternative and was successfully tested in human clinical trials. However, it is still challenging to achieve mechanical parameters comparative to permanent bare metal (BMS) and drug-eluting stents (DES). As such, in vitro tests are required to assess mechanical parameters correlated to the safety and efficacy of the device. Methods/Materials: In vitro bench tests evaluate scaffold profiles, length, deliverability, expansion behavior including acute elastic and time-dependent recoil, bending stiffness and radial strength. The Absorb GT1 (Abbott Vascular, Temecula, CA), DESolve (Elixir Medical Corporation, Sunnyvale, CA) and the Magmaris (BIOTRONIK AG, Bülach, Switzerland) that was previously tested in the BIOSOLVE II study, were tested. Results: Crimped profiles were 1.38 ± 0.01 mm (Absorb GT1), 1.39 ± 0.01 mm (DESolve) and 1.44 ± 0.00 mm (Magmaris) enabling 6F compatibility. Trackability was measured depending on stiffness and force transmission (pushability). Acute elastic recoil was measured at free expansion and within a mock vessel, respectively, yielding results of 5.86 ± 0.76 and 5.22 ± 0.38% (Absorb), 7.85 ± 3.45 and 9.42 ± 0.21% (DESolve) and 5.57 ± 0.72 and 4.94 ± 0.31% (Magmaris). Time-dependent recoil (after 1 h) was observed for the Absorb and DESolve scaffolds but not for the Magmaris. The self-correcting wall apposition behavior of the DESolve did not prevent time-dependent recoil under vessel loading. Conclusions: The results of the suggested test methods allow assessment of technical feasibility based on objective mechanical data and highlight the main differences between polymeric and metallic bioresorbable scaffolds.

  1. Flow cytometric investigations of diploid and tetraploid plants and in vitro cultures of Datura stramonium and Hyoscyamus niger.

    Science.gov (United States)

    Weber, Jost; Georgiev, Vasil; Pavlov, Atanas; Bley, Thomas

    2008-10-01

    Plant in vitro systems are valuable sources for the production of biological active substances. However, changed profiles of secondary metabolites, and low, variable yields possibly caused by genetic instabilities complicate their industrial implementation. DNA profiling of plant in vitro cultures may provide data for the selection of highly producing in vitro cultures. Diploid and tetraploid Datura stramonium and Hyoscyamus niger plant as well as calli, and hairy root lines derived from them were analyzed by flow cytometry. Plant in vitro cultures undergo several cycles of endoreduplication more than the explants from which they were obtained. The highest cycle values were observed in calli (e.g. 1.19 for diploid H. niger) possibly induced by the growth factors. However, hairy roots cultivated without growth factor exhibited significant degrees of endoreduplication (cycle value 0.88 for diploid H. niger). Sets of five hairy root lines from each plant and ploidy level showed consistent within-set ploidy patterns. The ploidy profiles of investigated plant in vitro and in vivo differ. For the first time we report that hairy roots of two Solanaceae species undergo endoreduplication. Theploidy profiles of in vitro cultures (hairy roots and calli) seem to be influenced by the genome size, the growth factors applied, and the type of in vitro culture. The transformation of several hairy root lines showed no differences in the ploidy patterns. Copyright 2008 International Society for Advancement of Cytometry.

  2. Raman spectroscopic approach to monitor the in vitro cyclization of creatine → creatinine

    Science.gov (United States)

    Gangopadhyay, Debraj; Sharma, Poornima; Singh, Sachin Kumar; Singh, Pushkar; Tarcea, Nicolae; Deckert, Volker; Popp, Jürgen; Singh, Ranjan K.

    2015-01-01

    The creatine → creatinine cyclization, an important metabolic phenomenon has been initiated in vitro at acidic pH and studied through Raman spectroscopic and DFT approach. The equilibrium composition of neutral, zwitterionic and protonated microspecies of creatine has been monitored with time as the reaction proceeds. Time series Raman spectra show clear signature of creatinine formation at pH 3 after ∼240 min at room temperature and reaction is faster at higher temperature. The spectra at pH 1 and pH 5 do not show such signature up to 270 min implying faster reaction rate at pH 3.

  3. In Vitro Investigation of Influences of Chitosan Nanoparticles on Fluorescein Permeation into Alveolar Macrophages.

    Science.gov (United States)

    Chachuli, Siti Haziyah Mohd; Nawaz, Asif; Shah, Kifayatullah; Naharudin, Idanawati; Wong, Tin Wui

    2016-06-01

    Pulmonary infection namely tuberculosis is characterized by alveolar macrophages harboring a large microbe population. The chitosan nanoparticles exhibit fast extracellular drug release in aqueous biological milieu. This study investigated the matrix effects of chitosan nanoparticles on extracellular drug diffusion into macrophages. Oligo, low, medium and high molecular weight chitosan nanoparticles were prepared by nanospray drying technique. These nanoparticles were incubated with alveolar macrophages in vitro and had model drug sodium fluorescein added into the same cell culture. The diffusion characteristics of sodium fluorescein and nanoparticle behavior were investigated using fluorescence microscopy, scanning electron microscopy, differential scanning calorimetry and Fourier transform infrared spectroscopy techniques. The oligochitosan nanoparticles enabled macrophage membrane fluidization with the extent of sodium fluorescein entry into macrophages being directly governed by the nanoparticle loading. Using nanoparticles made of higher molecular weight chitosan, sodium fluorescein permeation into macrophages was delayed due to viscous chitosan diffusion barrier at membrane boundary. Macrophage-chitosan nanoparticle interaction at membrane interface dictates drug migration into cellular domains.

  4. Synthesis and in vitro investigation of sol-gel derived bioglass-58S nanopowders

    Science.gov (United States)

    Joughehdoust, S.; Manafi, S.

    2012-03-01

    The aim of this research is the synthesis of bioglass-58S nanopowders by sol-gel method. Also, the effect of aging time of parent sols on the morphology, structure and particle size was investigated. Bioglass-58S powders were analyzed by X-ray diffraction patterns (XRD), Fourier transform infrared spectroscopy (FTIR), zetasizer instrument, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The XRD results showed that the powder is amorphous and glassy. According to FTIR spectroscopy, silicate bonds were formed in all powders. Zetasizer curves proved that the particle sizes of the powders and agglomerates have increased with aging time. The SEM images confirmed these results, too. Additionally, the TEM observations revealed that the increase of aging time caused the growth of grains with the size between 50-200 nm. The in vitro biological behavior of bioglass-58S powders were investigated by immersing the bioglass discs (made from the powders) in the simulated body fluid (SBF). The XRD patterns and SEM images confirmed the formation of the hydroxyapatite (HA) phase.

  5. Investigation of in vitro Opioid Receptor Binding Activities of Some Turkish Salvia species

    Directory of Open Access Journals (Sweden)

    Özge Gündüz Çınar

    2011-01-01

    Full Text Available Kappa Opioid Peptide Receptor (KOPr activation produces analgesic, psychotomimetic, diuretic and antipruritic effects. KOPr ligands are investigated for their potential roles in the treatment of addiction, depression, feeding behavior, psychosis and schizophrenia. In this study the methanolic extracts of a number of Salvia species which are native to Turkey (S. tomentosa, S. tchihatcheffii , S. rosifolia, S. dichroantha and S. sclarea were tested for their potential binding to opioid receptors in rat brain membranes and Chinese Hamster Ovary Cells expressing human KOPr (CHO-KOPh. [ 3H]Diprenorphine, an unselective opioid antagonist, was utilized in the radioligand receptor binding assays. All extracts (0.11 mg/ml inhibited the [ 3H]Diprenorphine binding with ranging KOPr binding affinities. More than 50% inhibition of diprenorphine binding was shown only with Salvia dichroantha and Salvia sclarea both in rat brain membranes and CHO-KOPh membranes.Among them Salvia sclarea deserves further investigation for its active component(s and its pharmacological characterization. This study clearly demonstrates the potential opioid receptor binding activities of several Turkish Salvia species. This work constitutes the first study on in vitro opioid receptor binding activities of Salvia species from the Turkish flora.

  6. A simplified approach for estimating skin permeation parameters from in vitro finite dose absorption studies.

    Science.gov (United States)

    Lehman, Paul A

    2014-12-01

    Historically, percutaneous absorption permeation parameters have been derived from in vitro infinite dose studies, yet there is uncertainty in their accuracy if the applied vehicle saturates or damages the stratum corneum, or when the permeation parameters are inappropriately derived from cumulative absorption data. An approach is provided for determining penetration parameters from in vitro finite dose data. Key variables, and equations for their derivation, are identified from the literature and provide permeation parameters that use only Tmax , AUC, and AUMC from finite dose data. The equations are tested with computer-generated model data and to actual study data. Derived permeation parameters obtained from the computer model data match those used in generating the simulated finite dose data. Parameters obtained from actual study data reasonably and acceptably model the penetration profile kinetics of the study data. From in vitro finite dose absorption data, three parameters can be obtained: the diffusion transit time (td ), which characterizes the diffusion coefficient, the partition volume (Vm P), which characterizes the partition coefficient, and the permeation coefficient (Kp ). These parameters can be obtained from finite dose data without having to know the length of the diffusion pathway through the membrane. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  7. An integrated approach for the in vitro dosimetry of engineered nanomaterials

    Science.gov (United States)

    2014-01-01

    Background There is a great need for screening tools capable of rapidly assessing nanomaterial toxicity. One impediment to the development of reliable in vitro screening methods is the need for accurate measures of cellular dose. We present here a methodology that enables accurate determination of delivered to cell dose metrics. This methodology includes (1) standardization of engineered nanomaterial (ENM) suspension preparation; (2) measurement of ENM characteristics controlling delivery to cells in culture; and (3) calculation of delivered dose as a function of exposure time using the ISDD model. The approach is validated against experimentally measured doses, and simplified analytical expressions for the delivered dose (Relevant In Vitro Dose (RID)f function) are derived for 20 ENMs. These functions can be used by nanotoxicologists to accurately calculate the total mass (RIDM), surface area (RIDSA), or particle number (RIDN) delivered to cells as a function of exposure time. Results The proposed methodology was used to derive the effective density, agglomerate diameter and RID functions for 17 industrially-relevant metal and metal oxide ENMs, two carbonaceous nanoparticles, and non-agglomerating gold nanospheres, for two well plate configurations (96 and 384 well plates). For agglomerating ENMs, the measured effective density was on average 60% below the material density. We report great variability in delivered dose metrics, with some materials depositing within 24 hours while others require over 100 hours for delivery to cells. A neutron-activated tracer particle system was employed to validate the proposed in vitro dosimetry methodology for a number of ENMs (measured delivered to cell dose within 9% of estimated). Conclusions Our findings confirm and extend experimental and computational evidence that agglomerate characteristics affect the dose delivered to cells. Therefore measurement of these characteristics is critical for effective use of in vitro systems

  8. An integrated approach for the in vitro dosimetry of engineered nanomaterials.

    Science.gov (United States)

    Cohen, Joel M; Teeguarden, Justin G; Demokritou, Philip

    2014-05-01

    There is a great need for screening tools capable of rapidly assessing nanomaterial toxicity. One impediment to the development of reliable in vitro screening methods is the need for accurate measures of cellular dose. We present here a methodology that enables accurate determination of delivered to cell dose metrics. This methodology includes (1) standardization of engineered nanomaterial (ENM) suspension preparation; (2) measurement of ENM characteristics controlling delivery to cells in culture; and (3) calculation of delivered dose as a function of exposure time using the ISDD model. The approach is validated against experimentally measured doses, and simplified analytical expressions for the delivered dose (Relevant In Vitro Dose (RID)f function) are derived for 20 ENMs. These functions can be used by nanotoxicologists to accurately calculate the total mass (RIDM), surface area (RIDSA), or particle number (RIDN) delivered to cells as a function of exposure time. The proposed methodology was used to derive the effective density, agglomerate diameter and RID functions for 17 industrially-relevant metal and metal oxide ENMs, two carbonaceous nanoparticles, and non-agglomerating gold nanospheres, for two well plate configurations (96 and 384 well plates). For agglomerating ENMs, the measured effective density was on average 60% below the material density. We report great variability in delivered dose metrics, with some materials depositing within 24 hours while others require over 100 hours for delivery to cells. A neutron-activated tracer particle system was employed to validate the proposed in vitro dosimetry methodology for a number of ENMs (measured delivered to cell dose within 9% of estimated). Our findings confirm and extend experimental and computational evidence that agglomerate characteristics affect the dose delivered to cells. Therefore measurement of these characteristics is critical for effective use of in vitro systems for nanotoxicology. The mixed

  9. Investigations on Actuator Dynamics through Theoretical and Finite Element Approach

    Directory of Open Access Journals (Sweden)

    Somashekhar S. Hiremath

    2010-01-01

    Full Text Available This paper gives a new approach for modeling the fluid-structure interaction of servovalve component-actuator. The analyzed valve is a precision flow control valve-jet pipe electrohydraulic servovalve. The positioning of an actuator depends upon the flow rate from control ports, in turn depends on the spool position. Theoretical investigation is made for No-load condition and Load condition for an actuator. These are used in finite element modeling of an actuator. The fluid-structure-interaction (FSI is established between the piston and the fluid cavities at the piston end. The fluid cavities were modeled with special purpose hydrostatic fluid elements while the piston is modeled with brick elements. The finite element method is used to simulate the variation of cavity pressure, cavity volume, mass flow rate, and the actuator velocity. The finite element analysis is extended to study the system's linearized response to harmonic excitation using direct solution steady-state dynamics. It was observed from the analysis that the natural frequency of the actuator depends upon the position of the piston in the cylinder. This is a close match with theoretical and simulation results. The effect of bulk modulus is also presented in the paper.

  10. Expert consensus on an in vitro approach to assess pulmonary fibrogenic potential of aerosolized nanomaterials.

    Science.gov (United States)

    Clippinger, Amy J; Ahluwalia, Arti; Allen, David; Bonner, James C; Casey, Warren; Castranova, Vincent; David, Raymond M; Halappanavar, Sabina; Hotchkiss, Jon A; Jarabek, Annie M; Maier, Monika; Polk, William; Rothen-Rutishauser, Barbara; Sayes, Christie M; Sayre, Phil; Sharma, Monita; Stone, Vicki

    2016-07-01

    The increasing use of multi-walled carbon nanotubes (MWCNTs) in consumer products and their potential to induce adverse lung effects following inhalation has lead to much interest in better understanding the hazard associated with these nanomaterials (NMs). While the current regulatory requirement for substances of concern, such as MWCNTs, in many jurisdictions is a 90-day rodent inhalation test, the monetary, ethical, and scientific concerns associated with this test led an international expert group to convene in Washington, DC, USA, to discuss alternative approaches to evaluate the inhalation toxicity of MWCNTs. Pulmonary fibrosis was identified as a key adverse outcome linked to MWCNT exposure, and recommendations were made on the design of an in vitro assay that is predictive of the fibrotic potential of MWCNTs. While fibrosis takes weeks or months to develop in vivo, an in vitro test system may more rapidly predict fibrogenic potential by monitoring pro-fibrotic mediators (e.g., cytokines and growth factors). Therefore, the workshop discussions focused on the necessary specifications related to the development and evaluation of such an in vitro system. Recommendations were made for designing a system using lung-relevant cells co-cultured at the air-liquid interface to assess the pro-fibrogenic potential of aerosolized MWCNTs, while considering human-relevant dosimetry and NM life cycle transformations. The workshop discussions provided the fundamental design components of an air-liquid interface in vitro test system that will be subsequently expanded to the development of an alternative testing strategy to predict pulmonary toxicity and to generate data that will enable effective risk assessment of NMs.

  11. In vitro investigation on the magnetic thermochemotherapy mediated by magnetic nanoparticles combined with methotrexate for breast cancer treatment.

    Science.gov (United States)

    Zhao, Lingyun; Huo, Meijun; Liu, Jiayi; Yao, Zhu; Li, Danye; Zhao, Zhiwei; Tang, Jintian

    2013-02-01

    Cancer comprehensive treatment has been fully recognized as it can provide an effective multimodality approach for fighting cancers. In therapeutic oncology, hyperthermic adjuvant chemotherapy termed as thermochemotherapy plays an increasing role in multimodality cancer treatment. Currently, targeted nanothermotherapy is one of the effective hyperthermia approach based on magnetic nanoparticles (MNPs), which can be achieved by applying biocompatible nanoscaled metallic particles that convert electromagnetic energy into heat, for instance, magnetic fluid hyperthermia (MFH) mediated by superparamagnetic iron oxide nanoparticles (SPIONs). Upon exposure under alternative magnetic field (AMF), SPIONs can generate heat through oscillation of their magnetic moment. Nowadays, clinical trials at phase II are now under investigations for MFH on patients in Germany and Japan and demonstrate very inspiring for cancer therapy. In this work we explore the feasibility and effectiveness of magnetic thermochemotherapy mediated by magnetic nanoparticles combined with methotrexate, an anti-cancer drug, for breast cancer comprehensive treatment. Amino silane coated MNPs as agent of MFH were prepared by the chemical precipitation method. Physiochemical characterizations on MNPs have been systematically carried out by various instrumental analyses. Inductive heating property of the MNPs was evaluated by monitoring the temperature increase of the MNPs suspension under AMF. The in-vitro cytotoxicity results on human breast cancer cell MCF-7 by CCK-8 assay indicated the bi-modal cancer treatment approach for combined MFH and chemotherapy is more effective than mono-modal treatment, indicating a thermal enhancement effect of hyperthermia on drug cytocoxicity. The magnetic thermochemotherapy mediated by MNPs combined with methotrexate can realize cancer comprehensive treatment thus has great potential in clinical application.

  12. A Novel Approach of Synthesizing and Evaluating the Anticancer Potential of Silver Oxide Nanoparticles in vitro.

    Science.gov (United States)

    Banerjee, Kaushik; Das, Satyajit; Choudhury, Pritha; Ghosh, Sarbari; Baral, Rathindranath; Choudhuri, Soumitra Kumar

    2017-01-01

    Development of novel strategies to kill cancer by sparing normal cells is of utmost importance. Apart from their known antimicrobial activity, only limited information has been recorded regarding the antitumor potential of biocompatible silver oxide nanoparticles (AgONPs). There is a need to evaluate the anticancer potential of biocompatible AgONPs in vitro. A new approach of utilizing the leaf extract of Excoecaria agallocha was used to synthesize AgONPs. This was then characterized by ultraviolet-visible spectrophotometry, nanoparticle-tracking analysis, and ζ-potential analysis. Cytotoxicity and apoptotic potential were evaluated with an MTT assay and an annexin V-binding assay against the murine melanoma (B16F10), murine colon cancer (CT26), murine lung adenocarcinoma (3LL), and murine Ehrlich ascites carcinoma (EAC) cell lines. Cellular localization of AgONPs was evaluated on fluorescence microscopy. UV peaks at 270 and 330 nm indicated the formation of nanoparticles (NPs) and the NP-tracking analyzer revealed them to have a size of 228 nm. AgONPs exerted initial cytotoxicity, specifically against all the experimental malignant cells by sparing the normal cell lines. Moreover, AgONPs exert apoptosis equally on all the malignant cells in vitro and ex vivo. This cytotoxicity possibly occurs via the nuclear translocation of AgONPs as analyzed in B16F10 cells. AgONPs utilizing natural sources would be a new medicinal approach against a broad spectrum of malignancy. © 2017 S. Karger AG, Basel.

  13. Investigation of subrosion processes using an integrated geophysical approach

    Science.gov (United States)

    Miensopust, Marion; Hupfer, Sarah; Kobe, Martin; Schneider-Löbens, Christiane; Wadas, Sonja

    2017-04-01

    Subrosion, i.e., leaching of readily soluble rocks, is usually of natural origin but can be enhanced by anthropogenic interferences. In recent years, public awareness of subrosion processes in terms of the in parts catastrophic implications and incidences increased. Especially the sinkholes in Schmalkalden, Tiefenort and Nordhausen (Germany) are three dramatic examples. They show that the knowledge of those processes and therefore, the predictability of such events is insufficient. The complexity of subrosion processes requires an integrated geophysical approach, which investigates the interlinking of structure, hydraulics, leaching, and mechanics. This contributes to a better understanding of the processes by reliable imaging and characterisation of subrosion structures. At LIAG an inter-sectional group is engaged in geophysical investigation of subrosion processes. The focus is application, enhancement and combination of various geophysical methods both at surface and in boreholes. This includes the monitoring of surface deformation and the application of time-lapse gravity as well as seismic, geoelectric and electromagnetic methods. Petrophysical investigations (with focus on Spectral Induced Polarisation - SIP) are conducted to characterise the processes on pore scale. Numerical studies are applied to advance the understanding of void forming processes and the mechanical consequences in the dynamic interaction. Since March 2014, quarterly campaigns are conducted to monitor changes in gravity acceleration at 15 stations in the urban area of Bad Frankenhausen. The standard deviations of the adjusted gravity differences are in the single-digit µGal range. The gravity acceleration changes in the range of 0 to 15 µGal over a timespan of three years and the accompanying levelling locally shows continuous subsidence in the mm/year-range. Sixteen SH-wave and four P-wave reflection seismic profiles together with three VSṔs were surveyed in the city of Bad

  14. Geophysical investigation of subrosion processes - an integrated approach

    Science.gov (United States)

    Miensopust, Marion; Hupfer, Sarah; Kobe, Martin; Schneider-Löbens, Christiane; Wadas, Sonja; Krawczyk, Charlotte

    2016-04-01

    Subrosion, i.e., leaching of readily soluble rocks mostly due to groundwater, is usually of natural origin but can be enhanced by anthropogenic interferences. In recent years, public awareness of subrosion processes in terms of the in parts catastrophic implications and incidences increased. Especially the sinkholes in Schmalkalden and Tiefenort (Germany) are - based on unforeseen collapse events and associated damage in 2010 - two dramatic examples. They illustrate that to date the knowledge of those processes and therefore the predictability of such events is insufficient. The complexity of the processes requires an integrated geophysical approach which investigates the interlinking of structure, hydraulics, solution processes, and mechanics. This finally contributes to a better understanding of the processes by reliable imaging and characterisation of subrosion structures. At LIAG an inter-sectional group is engaged in geophysical investigation of subrosion processes. The focus is application, enhancement and combination of various geophysical methods both at surface and in boreholes. This includes monitoring of (surface) deformation and variation of gravity as well as seismic, geoelectric and electromagnetic methods. Petrophysical investigations (with focus on spectral induced polarisation - SIP) are conducted to characterise the processes on pore scale. Numerical studies are applied to advance the understanding of void forming processes and the mechanical consequences in the dynamic interaction. Since March 2014, quarterly campaigns are conducted to monitor time-lapse gravity changes at 12 stations in the urban area of Bad Frankenhausen. The standard deviations of the gravity differences between the survey points are low and the accompanying levelling locally shows continuous subsidence in the mm/year-range. Eight shear-wave reflection seismic profiles were surveyed in Bad Frankenhausen using a landstreamer and an electro-dynamic vibrator. This method is

  15. Performance properties of the population bioequivalence approach for in vitro delivered dose for orally inhaled respiratory products.

    Science.gov (United States)

    Morgan, Beth; Strickland, Helen

    2014-01-01

    Regulatory agencies, industry, and academia have acknowledged that in vitro assessments serve a role in establishing bioequivalence for second-entry drug product approvals as well as innovator post-approval drug product changes. For orally inhaled respiratory products (OIPs), the issues of correctly analyzing in vitro data and interpreting the results within the broader context of therapeutic equivalence have garnered significant attention. One of the recommended statistical tests for in vitro data is the population bioequivalence method (PBE). The current literature for assessing the PBE statistical approach for in vitro data assumes a log normal distribution. This paper focuses on an assessment of that assumption for in vitro delivered dose. Concepts in development of a statistical model are presented. The PBE criterion and hypotheses are written for the case when data follows a normal distribution, rather than log normal. Results of a simulation study are reported, characterizing the performance of the PBE approach when data are expected to be normally distributed, rather than log normal. In these cases, decisions using the PBE approach are not consistent for the same absolute mean difference that the test product is from the reference product. A conclusion of inequivalency will occur more often if the test product dose is lower than the reference product for the same deviation from target. These features suggest that more research is needed for statistical equivalency approaches for in vitro data.

  16. Effect of glucose variability on pathways associated with glucotoxicity in diabetes: Evaluation of a novel in vitro experimental approach.

    Science.gov (United States)

    Kuricová, Katarína; Pácal, Lukáš; Šoupal, Jan; Prázný, Martin; Kaňková, Kateřina

    2016-04-01

    Glycaemic variability (GV) has been hypothesized to increase the risk of diabetes complications; however, results of clinical studies are contradictory. The effect of GV on cell phenotypes has been investigated in vitro showing that GV may have more deleterious effect on cells that high glucose itself. However, methodology used to study GV in vitro differs significantly between studies and does not reflect in vivo situation. Therefore we aimed to establish clinically relevant an in vitro experimental approach for the study of GV that reflects intra-day glucose fluctuations of subjects with type 1 diabetes mellitus (T1DM) and of healthy subjects and to test how low and high GV affect expression of genes that protects cells from hyperglycaemia-induced damage. Human umbilical vein endothelial cells (HUVEC) were cultured 24h in medium with different glucose profiles: high GV, low GV and GV of healthy subjects-profiles created according to CGM of T1DM patients and healthy subjects. These profiles were compared to commonly used 5.5 and 25mmol/l glucose concentrations. Gene expression was determined using quantitative PCR. Our results showed general down-regulation of enzymes that are involved in the protection against hyperglycaemia-induced intracellular changes in both low and high GV compared to normal glycaemia similarly to the decrease induced by continuous hyperglycaemia. Gene expressions did not differ between high and low GV. Our data indicate that GV may have similar or even greater effect than continuous hyperglycaemia on the expression of several genes relevant to pathogenesis of diabetes microvascular complications. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. Assessment of membrane stabilizing activity from honey. An in-vitro approach.

    Science.gov (United States)

    Manukumar, Honnayakanahalli Marichenne Gowda; Umesha, Sharanaiah

    2015-01-01

    esent study was conducted to evaluate Manoflora (MF), Polyflora (PF), Polyflora forest (PFf), and Processed (Pro) honey varieties to compare the in-vitro anti-inflammatory effects of aqueous honey samples in dose dependent manner. In-vitro anti-inflammatory activity was evaluated using membrane stabilization assay of RBCs at different aqueous honey concentrations. Material and method. The present investigation carried out for selected varieties of honey against erythrocytes exposed to both heat and hypotonic lyses and inhibition of membrane damage was compared to the standard drug acetylsalicylic acid. Results. Membrane damage was inhibited in both the model hemolysis of erythrocytes in vitro in a concentration dependent manner. Hypotonic solution inducing damage was inhibited by aqueous honey sample in ascending order ranged from 8.25% to 97.76% at 10 to 50 mg/ml and standard drug acetylsalicylic acid showing hemolysis protection 96.09% at 100 μg/ml concentration. In heat induced hemolysis model aqueous honey sample exhibited its protecting property during external stress condition in all samples ranged from 0.44% to 21.23% at 10 to 50 mg/ml and acetylsalicylic acid showed 39.38% at 100 μg/ml concentration. Among the variety PFf showed highest protecting nature for hypotonic solution induced lyses (97.76%) and heat induced hemolysis (21.23%) at 50 mg/ml respectively. Conclusion. With these investigations data conclude that the model exhibits marked anti-inflammatory effect. Future research is to be carried out to identify the molecules responsible in honey and its mechanism. present study was conducted to evaluate Manoflora (MF), Polyflora (PF), Polyflora forest (PFf), and Processed (Pro) honey varieties to compare the in-vitro anti-inflammatory effects of aqueous honey samples in dose dependent manner. In-vitro anti-inflammatory activity was evaluated using membrane stabilization assay of RBCs at different aqueous honey concentrations. Material and method. The

  18. Lipid digestion of protein stabilized emulsions investigated in a dynamic in vitro gastro-intestinal model system

    NARCIS (Netherlands)

    Helbig, A.; Silletti, E.; Aken, G.A. van; Oosterveld, A.; Minekus, M.; Hamer, R.J.; Gruppen, H.

    2013-01-01

    This study investigated the effect of gastric passage of protein stabilized emulsions, i.e., whey protein isolate (WPI) and lysozyme, under dynamic in vitro conditions on both the gastric and intestinal lipolysis. Emulsions were prepared at neutral pH to enable an opposite surface charge.

  19. Lipid Digestion of Protein Stabilized Emulsions Investigated in a Dynamic In Vitro Gastro-Intestinal Model System

    NARCIS (Netherlands)

    Helbig, A.; Silletti, E.; Aken, van G.A.; Oosterveld, A.; Minekus, M.; Hamer, R.J.; Gruppen, H.

    2013-01-01

    This study investigated the effect of gastric passage of protein stabilized emulsions, i.e., whey protein isolate (WPI) and lysozyme, under dynamic in vitro conditions on both the gastric and intestinal lipolysis. Emulsions were prepared at neutral pH to enable an opposite surface charge.

  20. An investigation of gamma-radiation sensitivity on in vitro study of Hordeum vulgare L.

    Directory of Open Access Journals (Sweden)

    Pinaki Chaudhuri

    2014-01-01

    Full Text Available Gamma-radiation sensitivity was studied on in vitro condition in Hordeum vulgare L. The variation of callusing response assessed with the increasing level of gamma-radiation treatment and regeneration delayed at higher dose level.

  1. An in vitro investigation of endocrine disrupting effects of the mycotoxin alternariol

    Energy Technology Data Exchange (ETDEWEB)

    Frizzell, Caroline [Institute for Global Food Security, School of Biological Sciences, Queen' s University Belfast, Northern Ireland (United Kingdom); Ndossi, Doreen [Section of Experimental Biomedicine, Norwegian School of Veterinary Science, Oslo (Norway); Sokoine University of Agriculture, Morogoro (Tanzania, United Republic of); Kalayou, Shewit [Section of Experimental Biomedicine, Norwegian School of Veterinary Science, Oslo (Norway); Mekelle University College of Veterinary Medicine, Mekelle (Ethiopia); Eriksen, Gunnar S. [Norwegian Veterinary Institute, Oslo (Norway); Verhaegen, Steven [Section of Experimental Biomedicine, Norwegian School of Veterinary Science, Oslo (Norway); Sørlie, Morten [Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Ås (Norway); Elliott, Christopher T. [Institute for Global Food Security, School of Biological Sciences, Queen' s University Belfast, Northern Ireland (United Kingdom); Ropstad, Erik [Section of Experimental Biomedicine, Norwegian School of Veterinary Science, Oslo (Norway); Connolly, Lisa, E-mail: l.connolly@qub.ac.uk [Institute for Global Food Security, School of Biological Sciences, Queen' s University Belfast, Northern Ireland (United Kingdom)

    2013-08-15

    Alternariol (AOH) is a mycotoxin commonly produced by Alternaria alternata on a wide range of foods. Few studies to date have been performed to evaluate the effects of AOH on endocrine activity. The present study makes use of in vitro mammalian cellular based assays and gene expression to investigate the ability of AOH to act as an endocrine disruptor by various modes of action. Reporter gene assays (RGAs), incorporating natural steroid hormone receptors for oestrogens, androgens, progestagens and glucocorticoids were used to identify endocrine disruption at the level of nuclear receptor transcriptional activity, and the H295R steroidogenesis assay was used to assess endocrine disruption at the level of gene expression and steroid hormone production. AOH exhibited a weak oestrogenic response when tested in the oestrogen responsive RGA and binding of progesterone to the progestagen receptor was shown to be synergistically increased in the presence of AOH. H295R cells when exposed to 0.1–1000 ng/ml AOH, did not cause a significant change in testosterone and cortisol hormones but exposure to 1000 ng/ml (3.87 μM) AOH resulted in a significant increase in estradiol and progesterone production. In the gene expression study following exposure to 1000 ng/ml (3.87 μM) AOH, only one gene NR0B1 was down-regulated, whereas expression of mRNA for CYP1A1, MC2R, HSD3B2, CYP17, CYP21, CYP11B2 and CYP19 was up-regulated. Expression of the other genes investigated did not change significantly. In conclusion AOH is a weak oestrogenic mycotoxin that also has the ability to interfere with the steroidogenesis pathway. - Highlights: • Alternariol was investigated for endocrine disrupting activity. • Reporter gene assays and the H295R steroidogenesis assay have been used. • An oestrogenic effect of alternariol was observed. • This can lead to an increase in expression of the progesterone receptor. • Alternariol is capable of modulating hormone production and gene expression.

  2. Selection of a T7 promoter mutant with enhanced in vitro activity by a novel multi-copy bead display approach for in vitro evolution.

    Science.gov (United States)

    Paul, Siddhartha; Stang, Alexander; Lennartz, Klaus; Tenbusch, Matthias; Überla, Klaus

    2013-01-07

    In vitro evolution of nucleic acids and proteins is a powerful strategy to optimize their biological and physical properties. To select proteins with the desired phenotype from large gene libraries, the proteins need to be linked to the gene they are encoded by. To facilitate selection of the desired phenotype and isolation of the encoding DNA, a novel bead display approach was developed, in which each member of a library of beads is first linked to multiple copies of a clonal gene variant by emulsion polymerase chain reaction. Beads are transferred to a second emulsion for an in vitro transcription-translation reaction, in which the protein encoded by each bead's amplicon covalently binds to the bead present in the same picoliter reactor. The beads then contain multiple copies of a clonal gene variant and multiple molecules of the protein encoded by the bead's gene variant and serve as the unit of selection. As a proof of concept, we screened a randomized library of the T7 promoter for high expression levels by flow cytometry and identified a T7 promoter variant with an ~10-fold higher in vitro transcriptional activity, confirming that the multi-copy bead display approach can be efficiently applied to in vitro evolution.

  3. In vitro investigation of ultrasound-induced oxidative stress on human lens epithelial cells.

    Science.gov (United States)

    Rwei, Patrick; Alex Gong, Cihun-Siyong; Luo, Li-Jyuan; Lin, Meng-Bo; Lai, Jui-Yang; Liu, Hao-Li

    2017-01-22

    The effect of ultrasound exposure on human lens epithelial cells (HLE-B3) was investigated in vitro, specifically on the generation of oxidative stress upon ultrasound application using various clinically-relevant settings. In addition to ultrasound-induced heat effects, oxidative stress has been recently proposed as one of the main mechanisms for ultrasound-induced effects on human cells. In this work, the levels of biocompatibility and generation of oxidative stress by exposure of ultrasound to HLE-B3 were evaluated quantitatively and qualitatively by the MTT assay, Live/Dead assay, reactive oxygen species (ROS) and intracellular calcium level. Oxidative stress induction is traditionally achieved through administrations of H2O2 and thus the administration of H2O2 was used as the positive control group for comparison herein. Concerning the administrations of H2O2 are considered invasive and may potentially have side effects, ultrasound as physical stimulation could be a safer and non-invasive method to induce similar oxidative stress environments. The effect of ultrasound on cell viability and induction of oxidative stress increases with ultrasound intensity. The result reveals that the continuous ultrasound has a positive impact on the oxidative stress levels but does negatively on the cell viability, as compared to the pulsed ultrasound. Furthermore, our work demonstrates that the exposure of 58 kPa continuous ultrasound without microbubbles can maintain acceptable cell viability and produce oxidative stress effects similar to the traditional administrations of H2O2. In summary, exposure of ultrasound can generate oxidative stress comparable to traditional administrations of H2O2. The effect of generating oxidative stress is adjustable through ultrasound parameters, including the pulsed or continuous wave, the intensity of ultrasound and addition of microbubbles. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Proximal contact tightness between direct-composite additions in the posterior dentition: an in vitro investigation.

    Science.gov (United States)

    Wolff, Diana; Hahn, Philipp; Ding, Paul; Maier-Kraus, Theresa; Frese, Cornelia; Doerfer, Christof; Staehle, Hans Joerg

    2012-01-01

    The aim of the study was to test whether a novel three-step matrix technique for posterior direct-composite additions creates sufficiently strong proximal contacts. Contact tightness was measured between direct-composite additions and between original teeth on a model. Therefore, the frictional forces required to remove a straight, 0.05-mm-thick, metal matrix band inserted between adjacent teeth and held by a universal testing machine (Zwicki, Zwick GmbH, Ulm, Germany) were recorded. Measurements were taken at three time points to carry out reference analysis: at baseline, after removal of the maxillary right second premolar (tooth #15) to simulate a diastema, and after closure of the diastema by inserting two direct-composite additions with the three-step matrix technique on the maxillary right first premolar (tooth #14) and first molar (tooth #16). Measurements were performed in the maxillary right (first) and left (second) quadrants to document sagittal displacement. The original contact tightness values were between 1.65 ± 0.88 N and 3.05 ± 0.60 N in the first quadrant and between 1.23 ± 0.51 N and 2.18 ± 0.43 N in the second quadrant. After removal of tooth 15, values decreased significantly in the first quadrant and insignificantly in the second. After reconstruction, the contact tightness between teeth 14 and 16 was significantly stronger (tighter) (3.20 ± 0.80 N) than the originally measured contact tightness between teeth 14 and 15 (2.86 ± 0.64 N) and teeth 15 and 16 (1.65 ± 0.88 N) (p=0.006 and 0.001, respectively). Within the limitations of an in vitro investigation, this study has shown that by using a novel, three-step matrix technique, direct posterior composite additions can form sufficiently tight proximal contacts.

  5. Analytical in vitro approach for studying cyto- and genotoxic effects of particulate airborne material.

    Science.gov (United States)

    Aufderheide, Michaela; Scheffler, Stefanie; Möhle, Niklas; Halter, Beat; Hochrainer, Dieter

    2011-12-01

    In the field of inhalation toxicology, progress in the development of in vitro methods and efficient exposure strategies now offers the implementation of cellular-based systems. These can be used to analyze the hazardous potency of airborne substances like gases, particles, and complex mixtures (combustion products). In addition, the regulatory authorities require the integration of such approaches to reduce or replace animal experiments. Although the animal experiment currently still has to provide the last proof of the toxicological potency and classification of a certain compound, in vitro testing is gaining more and more importance in toxicological considerations. This paper gives a brief characterization of the CULTEX® Radial Flow System exposure device, which allows the exposure of cultivated cells as well as bacteria under reproducible and stable conditions for studying cellular and genotoxic effects after the exposure at the air-liquid or air-agar interface, respectively. A commercial bronchial epithelial cell line (16HBE14o-) as well as Salmonella typhimurium tester strains were exposed to smoke of different research and commercial available cigarettes. A dose-dependent reduction of cell viability was found in the case of 16HBE14o- cells; S. typhimurium responded with a dose-dependent induction of revertants. The promising results recommend the integration of cellular studies in the field of inhalation toxicology and their regulatory acceptance by advancing appropriate validation studies.

  6. A combined approach to early detect in vitro drug-induced hemostatic changes in preclinical safety.

    Science.gov (United States)

    Defontis, Myriam; Côté, Serge; Ledieu, David

    2017-06-14

    Early detection of drug-induced alterations of hemostasis is challenging. Drugs can affect different components of the Virchow's triad and measurement of plasmatic coagulation times lacks sensitivity. New techniques for a more global assessment of the hemostasis are now available: the impedance platelet aggregometry, the thromboelastography and the thrombin generation measurement. The aim of this study was to evaluate three techniques (i.e.: Multiplate®, TEG® and CAT) for the in vitro detection of the effect of a drug known to induce hemostatic alterations in a preclinical safety environment. Cyclosporine A was chosen and tested at 4 concentrations after solubilization in DMSO in Wistar rats and Beagle dogs. The results obtained were comparable between both species except for the thrombin generation in platelet rich plasma. Enhanced platelet aggregability was observed after ADP stimulation and alterations of the thromboelastograms consisted in decreased maximum amplitude and increased LY30. A dual effect on thrombin generation was observed and suggested that CsA may interact with platelets in rat platelet rich plasma and speed up thrombin generation. The results of this study indicate that using a combined approach on hemostasis testing in preclinical safety it is possible to detect in vitro drug-induced alterations of hemostasis. Copyright © 2017. Published by Elsevier GmbH.

  7. A combined in vitro approach to improve the prediction of mitochondrial toxicants.

    Science.gov (United States)

    Eakins, Julie; Bauch, Caroline; Woodhouse, Heather; Park, Benjamin; Bevan, Samantha; Dilworth, Clive; Walker, Paul

    2016-08-01

    Drug induced mitochondrial dysfunction has been implicated in organ toxicity and the withdrawal of drugs or black box warnings limiting their use. The development of highly specific and sensitive in vitro assays in early drug development would assist in detecting compounds which affect mitochondrial function. Here we report the combination of two in vitro assays for the detection of drug induced mitochondrial toxicity. The first assay measures cytotoxicity after 24h incubation of test compound in either glucose or galactose conditioned media (Glu/Gal assay). Compounds with a greater than 3-fold toxicity in galactose media compared to glucose media imply mitochondrial toxicity. The second assay measures mitochondrial respiration, glycolysis and a reserve capacity with mechanistic responses observed within one hour following exposure to test compound. In order to assess these assays a total of 72 known drugs and chemicals were used. Dose-response data was normalised to 100× Cmax giving a specificity, sensitivity and accuracy of 100%, 81% and 92% respectively for this combined approach. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. Investigations of the Biological Effects of Airborne and Inhalable Substances by Cell-Based In Vitro Methods: Fundamental Improvements to the ALI Concept

    Directory of Open Access Journals (Sweden)

    Detlef Ritter

    2014-01-01

    Full Text Available The state of the art for cell-based in vitro investigations of airborne and inhalable material is “air-liquid interface” (ALI technology. Cell lines, primary cells, complex 3D models, or precision-cut lung slices (PCLS are used to represent the lung or skin by way of an in vitro barrier model. These models have been applied in toxicity or pharmacological testing. However, contrasting with a clear demand for alternative methods, there is still no widely accepted procedure for cell-based in vitro testing of inhalable substances. In the light of this, an analysis was undertaken of common drawbacks of current approaches. Hence, the pivotal improvements aimed at were the cellular exposure environment, overall performance and applicability, operability of online investigations during exposure and routine setup. It resulted in an improved device (P.R.I.T. ExpoCube based on an “all-in-one-plate” concept including all phases of the experiment (cell culture, exposure, and read-out and all experimental groups (two test gas groups, controls in one single commercial multiwell plate. Verification of the concept was demonstrated in a first experimental series using reference substances (formaldehyde, ozone, and clean air. The resulting ALI procedure enables the application of inhalable substances and mixtures under highly effective exposure conditions in routine utilization.

  9. Introduction to social media investigation a hands-on approach

    CERN Document Server

    Golbeck, Jennifer

    2015-01-01

    If you're interested in using social media as an investigative tool, Introduction to Social Media Investigation will show you how! Social networks and social media, like Facebook, Twitter, and Foursquare, are some of the most popular services on the Web, with hundreds of millions of users. The public information that people share on these sites can be valuable for anyone interested in investigating people of interest through open, public sources. Social media as an investigative device is in its infancy and not well understood. This book presents an overview of social media and discusses s

  10. Can magnetic resonance spectroscopy differentiate malignant and benign causes of lymphadenopathy? An in-vitro approach.

    Science.gov (United States)

    Buré, Lionel; Boucher, Louis-Martin; Blumenkrantz, Miriam; Schob, Stefan; Lafaye de Micheaux, Pierre; Reinhold, Caroline; Gallix, Benoit

    2017-01-01

    Lymphadenopathy continues to be a common problem to radiologists and treating physicians because of the difficulty in confidently categorizing a node as being benign or malignant using standard diagnostic techniques. The goal of our research was to assess whether magnetic resonance (MR) spectroscopy contains the necessary information to allow differentiation of benign from malignant lymph nodes in an in-vitro approach using a modern pattern recognition method. Tissue samples from a tissue bank were analyzed on a nuclear magnetic resonance (NMR) spectrometer. A total of 69 samples were studied. The samples included a wide variety of malignant and benign etiologies. Using 45 samples, we initially created a model which was able to predict if a certain spectrum originates from benign or malignant lymph nodes using a pattern-recognition technique which takes into account the entire magnetic spectrum rather than single peaks alone. The remaining 24 samples were blindly loaded in the model to assess its performance. We obtained an excellent accuracy in differentiating benign and malignant lymphadenopathy using the model. It correctly differentiated as malignant or benign, in a blinded fashion, all of the malignant samples (13 of 13) and 10 out of the 11 benign samples. We thus showed that magnetic spectroscopy is able to differentiate benign from malignant causes of lymphadenopathy. Additional experiments were performed to verify that the differentiating abilities of our model were not due to differential tissue decay in between benign and malignant tissues. If future experiments demonstrate that a similar approach could be executed with standard MR imaging, this technique could be useful as a problem-solving tool when assessing lymphadenopathy in general. Alternatively, our in-vitro technique could also be useful to pathologists faced with indeterminate pathologies of the lymph nodes after validating our results with a larger sample size.

  11. An Investigative, Cooperative Learning Approach to the General Microbiology Laboratory

    Science.gov (United States)

    Seifert, Kyle; Fenster, Amy; Dilts, Judith A.; Temple, Louise

    2009-01-01

    Investigative- and cooperative-based learning strategies have been used effectively in a variety of classrooms to enhance student learning and engagement. In the General Microbiology laboratory for juniors and seniors at James Madison University, these strategies were combined to make a semester-long, investigative, cooperative learning experience…

  12. Enzyme Assay: An Investigative Approach to Enhance Science Process Skills

    Science.gov (United States)

    Vartak, Rekha; Ronad, Anupama; Ghanekar, Vikrant

    2013-01-01

    Scientific investigations play a vital role in teaching and learning the process of science. An investigative task that was developed for pre-university students is described here. The task involves extraction of an enzyme from a vegetable source and its detection by biochemical method. At the beginning of the experiment, a hypothesis is presented…

  13. Phytochemical investigation of natural and in vitro raised Vṛddhadāruka plants

    Directory of Open Access Journals (Sweden)

    Asha Jyoti Bharati

    2014-01-01

    Full Text Available Background: Argyreia nervosa commonly known as elephant creeper (English and Vṛddhadāruka (Sanskrit is a woody climber that belongs to the family Convolvulaceae. Seeds of this plant contain hallucinogens including ergot alkaloids and a naturally occurring lysergic acid amide. Traditionally the plant is used in the treatment of gonorrhea, strangury, chronic ulcers, diabetes, anemia and cerebral disorders. The plant is also used as appetitiser, brain tonic, cardiotonic, aphrodisiac. It possesses anti-inflammatory, immunomodulatory, antibacterial, antiviral and antifungal activities. Objective: To give an account of information on in vitro regeneration and phytochemical analysis of the plant. Materials and Methods: Nodal explants were selected for in vitro regeneration. Different aerial parts viz., seeds, natural and in vitro leaf, stem and callus were dried and extracted with different solvents and were subjected to various phytochemical analyses. Results: Different concentrations of 6-benzylaminopurine showed shoot and root initiation. The study of phytochemical screening of different extracts showed the presence of bioactive substances like flavonoids, alkaloids, terpenoids, etc. Conclusion: The study will provide an efficient in vitro protocol for micropropagation as an alternative method to conserve the plant and shows the presence of some important secondary metabolites in the nature grown and in vitro raised plants which can be useful for treatment of various diseases.

  14. Phytochemical investigation of natural and in vitro raised Vṛddhadāruka plants.

    Science.gov (United States)

    Bharati, Asha Jyoti; Bansal, Yogendra Kumar

    2014-01-01

    Argyreia nervosa commonly known as elephant creeper (English) and Vṛddhadāruka (Sanskrit) is a woody climber that belongs to the family Convolvulaceae. Seeds of this plant contain hallucinogens including ergot alkaloids and a naturally occurring lysergic acid amide. Traditionally the plant is used in the treatment of gonorrhea, strangury, chronic ulcers, diabetes, anemia and cerebral disorders. The plant is also used as appetitiser, brain tonic, cardiotonic, aphrodisiac. It possesses anti-inflammatory, immunomodulatory, antibacterial, antiviral and antifungal activities. To give an account of information on in vitro regeneration and phytochemical analysis of the plant. Nodal explants were selected for in vitro regeneration. Different aerial parts viz., seeds, natural and in vitro leaf, stem and callus were dried and extracted with different solvents and were subjected to various phytochemical analyses. Different concentrations of 6-benzylaminopurine showed shoot and root initiation. The study of phytochemical screening of different extracts showed the presence of bioactive substances like flavonoids, alkaloids, terpenoids, etc. The study will provide an efficient in vitro protocol for micropropagation as an alternative method to conserve the plant and shows the presence of some important secondary metabolites in the nature grown and in vitro raised plants which can be useful for treatment of various diseases.

  15. Investigation of In Vitro Bone Cell Adhesion and Proliferation on Ti Using Direct Current Stimulation.

    Science.gov (United States)

    Bodhak, Subhadip; Bose, Susmita; Kinsel, William C; Bandyopadhyay, Amit

    2012-12-01

    Our objective was to establish an in vitro cell culture protocol to improve bone cell attachment and proliferation on Ti substrate using direct current stimulation. For this purpose, a custom made electrical stimulator was developed and a varying range of direct currents, from 5 to 25 µA, were used to study the current stimulation effect on bone cells cultured on conducting Ti samples in vitro. Cell-materials interaction was studied for a maximum of 5 days by culturing with human fetal osteoblast cells (hFOB). The direct current was applied in every 8 h time interval and the duration of electrical stimulation was kept constant at 15 min for all cases. In vitro results showed that direct current stimulation significantly favored bone cell attachment and proliferation in comparison to nonstimulated Ti surface. Immunochemistry and confocal microscopy results confirmed that the cell adhesion was most pronounced on 25 µA direct current stimulated Ti surfaces as hFOB cells expressed higher vinculin protein with increasing amount of direct current. Furthermore, MTT assay results established that cells grew 30% higher in number under 25 µA electrical stimulation as compared to nonstimulated Ti surface after 5 days of culture period. In this work we have successfully established a simple and cost effective in vitro protocol offering easy and rapid analysis of bone cell-materials interaction which can be used in promotion of bone cell attachment and growth on Ti substrate using direct current electrical stimulation in an in vitro model.

  16. Investigating Approaches to Achieve Modularity Benefits in the Acquisition Ecosystem

    Science.gov (United States)

    2017-06-09

    Some additional questions that guided the research include:  Ecosystem concept : Based on a thorough literature review, what might be the “minimum...Summary of RT-163 Achievements ........................................................................................ 3 Phase A: Deep Dive Literature ...and explored the concept of a “MOSA ecosystem” that facilitates the adoption of strategies to achieve the desired benefits from a MOSA approach. The

  17. A sequential mixed methods research approach to investigating HIV ...

    African Journals Online (AJOL)

    2016-09-03

    Sep 3, 2016 ... show the effectiveness of the research method. The approach commenced with a quantitative survey to establish base-line information. Semi-structured interviews then yielded richer qualitative data before quantitative datasets were revisited to exploit their potential for more complex analysis and modelling ...

  18. Investigating the shape of the EKC: A nonparametric approach

    OpenAIRE

    Di Falco, Salvatore; Baiocchi, Giovanni

    2001-01-01

    In this paper we investigate the existence and shape of the Environmental Kuznets Curve (EKC) by means of nonparametric methods. We also investigate the issues involved in the choice of nonparametric estimator. We find that the nature of the economic relationship and the quality of environmental data can considerably impact estimates and therefore the implied policy recommendations. The flexible nature of nonparametric estimation allows us to develop a nonparametric test in the spirit of Silv...

  19. Data-driven approaches in the investigation of social perception.

    Science.gov (United States)

    Adolphs, Ralph; Nummenmaa, Lauri; Todorov, Alexander; Haxby, James V

    2016-05-05

    The complexity of social perception poses a challenge to traditional approaches to understand its psychological and neurobiological underpinnings. Data-driven methods are particularly well suited to tackling the often high-dimensional nature of stimulus spaces and of neural representations that characterize social perception. Such methods are more exploratory, capitalize on rich and large datasets, and attempt to discover patterns often without strict hypothesis testing. We present four case studies here: behavioural studies on face judgements, two neuroimaging studies of movies, and eyetracking studies in autism. We conclude with suggestions for particular topics that seem ripe for data-driven approaches, as well as caveats and limitations. © 2016 The Author(s).

  20. An investigative, cooperative learning approach to the general microbiology laboratory.

    Science.gov (United States)

    Seifert, Kyle; Fenster, Amy; Dilts, Judith A; Temple, Louise

    2009-01-01

    Investigative- and cooperative-based learning strategies have been used effectively in a variety of classrooms to enhance student learning and engagement. In the General Microbiology laboratory for juniors and seniors at James Madison University, these strategies were combined to make a semester-long, investigative, cooperative learning experience involving culture and identification of microbial isolates that the students obtained from various environments. To assess whether this strategy was successful, students were asked to complete a survey at the beginning and at the end of the semester regarding their comfort level with a variety of topics. For most of the topics queried, the students reported that their comfort had increased significantly during the semester. Furthermore, this group of students thought that the quality of this investigative lab experience was much better than that of any of their previous lab experiences.

  1. A Simple "in Vitro" Culture of Freshwater Prawn Embryos for Laboratory Investigations

    Science.gov (United States)

    Porntrai, Supaporn; Damrongphol, Praneet

    2008-01-01

    Giant freshwater prawn ("Macrobrachium rosenbergii" De Man) embryos can be cultured "in vitro" to hatching in 15% (v/v) artificial seawater (ASW). This technique can be applied as a bioassay for testing toxicity or for the effects of various substances on embryo development and can be used as a simple and low-cost model for…

  2. A New Phylogenetic Approach to Investigate the Intestinal Bacterial Composition

    DEFF Research Database (Denmark)

    Hermann-Bank, Marie Louise; Mølbak, Lars

    sequencing. The objective of this study is to demonstrate the applicability of the Access Array 48.48 (AA48.48) from Fluidigm® in investigating the composition of the intestinal microbiota. The array makes it possible to analyse 48 primer pairs against 48 samples resulting in 2304 individual Real-Time PCR...

  3. A Multimethod Approach for Investigating Algal Toxicity of Platinum Nanoparticles

    DEFF Research Database (Denmark)

    Sørensen, Sara Nørgaard; Engelbrekt, Christian; Lützhøft, Hans-Christian Holten

    2016-01-01

    The ecotoxicity of platinum nanoparticles (PtNPs) widely used in for example automotive catalytic converters, is largely unknown. This study employs various characterization techniques and toxicity end points to investigate PtNP toxicity toward the green microalgae Pseudokirchneriella subcapitata...

  4. Investigating the Development of Foreign Language Anxiety: An Autobiographical Approach

    Science.gov (United States)

    Trang, Tran Thi Thu; Baldauf, Richard B., Jr.; Moni, Karen

    2013-01-01

    Foreign language anxiety (FLA) has been found to exist in tertiary students learning foreign languages in many countries; however, limited research has explored how it develops. This study investigated how anxiety developed in students of English as a foreign language (EFL), focusing on changes in their feelings about EFL learning as they learned,…

  5. Establishing in vitro-in vivo correlation for antibody drug conjugate efficacy: a PK/PD modeling approach.

    Science.gov (United States)

    Shah, Dhaval K; Loganzo, Frank; Haddish-Berhane, Nahor; Musto, Sylvia; Wald, Hallie S; Barletta, Frank; Lucas, Judy; Clark, Tracey; Hansel, Steve; Betts, Alison

    2018-02-08

    The objective of this manuscript was to establish in vitro-in vivo correlation (IVIVC) between the in vitro efficacy and in vivo efficacy of antibody drug conjugates (ADCs), using a PK/PD modeling approach. Nineteen different ADCs were used to develop IVIVC. In vitro efficacy of ADCs was evaluated using a kinetic cell cytotoxicity assay. The cytotoxicity data obtained from in vitro studies was characterized using a novel mathematical model, parameter estimates from which were used to derive an in vitro efficacy matrix for each ADC, termed as 'in vitro tumor static concentration' (TSCin vitro). TSCin vitro is a theoretical concentration at continuous exposure of which the number of cells will neither increase nor decrease, compared to the initial cell number in the experiment. The in vivo efficacy of ADCs was evaluated using tumor growth inhibition (TGI) studies performed on human tumor xenograft bearing mice. The TGI data obtained from in vivo studies was characterized using a PK/PD model, parameter estimates from which were used to derive an in vivo efficacy matrix for each ADC, termed as 'in vivo tumor static concentration' (TSCin vivo). TSCin vivo is a theoretical concentration if one were to maintain in the plasma of a tumor bearing mouse, the tumor volume will neither increase nor decrease compared to the initial tumor volume. Comparison of the TSCin vitro and TSCin vivo values from 19 ADCs provided a linear and positive IVIVC. The Spearman's rank correlation coefficient for TSCin vitro and TSCin vivo was found to be 0.82. On average TSCin vivo was found to be ~ 27 times higher than TSCin vitro. The reasonable IVIVC for ADCs suggests that in vitro efficacy data was correctly able to differentiate ADCs for their in vivo efficacy. Thus, IVIVC can be used as a tool to triage ADC molecules in the discovery stage, thereby preventing unnecessary scaling-up of ADCs and waste of time and resources. An ability to predict the concentration of ADC that is efficacious

  6. Nest expansion assay: a cancer systems biology approach to in vitro invasion measurements

    Directory of Open Access Journals (Sweden)

    Estrada Lourdes

    2009-07-01

    Full Text Available Abstract Background Traditional in vitro cell invasion assays focus on measuring one cell parameter at a time and are often less than ideal in terms of reproducibility and quantification. Further, many techniques are not suitable for quantifying the advancing margin of collectively migrating cells, arguably the most important area of activity during tumor invasion. We have developed and applied a highly quantitative, standardized, reproducible Nest Expansion Assay (NEA to measure cancer cell invasion in vitro, which builds upon established wound-healing techniques. This assay involves creating uniform circular "nests" of cells within a monolayer of cells using a stabilized, silicone-tipped drill press, and quantifying the margin expansion into an overlaid extracellular matrix (ECM-like component using computer-assisted applications. Findings The NEA was applied to two human-derived breast cell lines, MCF10A and MCF10A-CA1d, which exhibit opposite degrees of tumorigenicity and invasion in vivo. Assays were performed to incorporate various microenvironmental conditions, in order to test their influence on cell behavior and measures. Two types of computer-driven image analysis were performed using Java's freely available ImageJ software and its FracLac plugin to capture nest expansion and fractal dimension, respectively – which are both taken as indicators of invasiveness. Both analyses confirmed that the NEA is highly reproducible, and that the ECM component is key in defining invasive cell behavior. Interestingly, both analyses also detected significant differences between non-invasive and invasive cell lines, across various microenvironments, and over time. Conclusion The spatial nature of the NEA makes its outcome susceptible to the global influence of many cellular parameters at once (e.g., motility, protease secretion, cell-cell adhesion. We propose the NEA as a mid-throughput technique for screening and simultaneous examination of factors

  7. Efficient and simple approach to in vitro culture of primary epithelial cancer cells.

    Science.gov (United States)

    Janik, Karolina; Popeda, Marta; Peciak, Joanna; Rosiak, Kamila; Smolarz, Maciej; Treda, Cezary; Rieske, Piotr; Stoczynska-Fidelus, Ewelina; Ksiazkiewicz, Magdalena

    2016-12-01

    Primary cancer cells constitute a favourable testing platform for in vitro research in oncology field as they reflect tumour state more accurately than the most commonly employed stable cell lines. Unfortunately, due to limited availability of material and difficulties with protocols validation, primary models are rarely implemented into laboratory practice.We have compared protocols for primary cultures, differing in media components and plate coatings. In terms of culture establishment, application of Geltrex® coating demonstrated equal efficiency to feeder layer (83% compared with 72% successfully established breast and 80% compared with 80% prostate tumour specimens), yet it was substantially less complicated and easier to validate. Both Geltrex® coating and tissue-specific primary cell medium were permanently required to successfully maintain primary epithelial prostate cancer cells (PEPCs) in culture. In case of primary epithelial breast cancer cells (PEBCs), collagen I coating enabled to obtain comparable number of passages to Geltrex® coating (P=0.438). Commercial primary cell media demonstrated lower efficiency than tissue-specific ones (PEPCs-5 compared with 8 and PEBCs-6 compared with 9 passages). Interestingly, both analysed tumour types were unsusceptible to induction of culture lifespan extension when transduced with SV40LT, BMI-1 or hEST2 genes, commonly applied as potential immortalizing agents.In conclusion, the approach based on extracellular matrix reconstitution and tissue-specific primary cell media is easy to validate and provides in vitro expansion sufficient for analytical purposes (approximately 8 passages). Therefore, it may facilitate implementation of hardly available experimental models for a variety of analyses. © 2016 The Author(s).

  8. A Support Vector Regression Approach for Investigating Multianticipative Driving Behavior

    Directory of Open Access Journals (Sweden)

    Bin Lu

    2015-01-01

    Full Text Available This paper presents a Support Vector Regression (SVR approach that can be applied to predict the multianticipative driving behavior using vehicle trajectory data. Building upon the SVR approach, a multianticipative car-following model is developed and enhanced in learning speed and predication accuracy. The model training and validation are conducted by using the field trajectory data extracted from the Next Generation Simulation (NGSIM project. During the model training and validation tests, the estimation results show that the SVR model performs as well as IDM model with respect to the model prediction accuracy. In addition, this paper performs a relative importance analysis to quantify the multianticipation in terms of the different stimuli to which drivers react in platoon car following. The analysis results confirm that drivers respond to the behavior of not only the immediate leading vehicle in front but also the second, third, and even fourth leading vehicles. Specifically, in congested traffic conditions, drivers are observed to be more sensitive to the relative speed than to the gap. These findings provide insight into multianticipative driving behavior and illustrate the necessity of taking into account multianticipative car-following model in microscopic traffic simulation.

  9. Investigation of a spatial double pendulum: an engineering approach

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available The behavior of a spatial double pendulum (SDP, comprising two pendulums that swing in different planes, was investigated. Movement equations (i.e., mathematical model were derived for this SDP, and oscillations of the system were computed and compared with experimental results. Matlab computer programs were used for solving the nonlinear differential equations by the Runge-Kutta method. Fourier transformation was used to obtain the frequency spectra for analyses of the oscillations of the two pendulums. Solutions for free oscillations of the pendulums and graphic descriptions of changes in the frequency spectra were used for the dynamic investigation of the pendulums for different initial conditions of motion. The value of the friction constant was estimated experimentally and incorporated into the equations of motion of the pendulums. This step facilitated the comparison between the computed and measured oscillations.

  10. Ultrasound - A new approach for non-woven scaffolds investigation

    Science.gov (United States)

    Khramtsova, E. A.; Morokov, E. S.; Lukanina, K. I.; Grigoriev, T. E.; Petronyuk, Y. S.; Levin, V. M.

    2016-05-01

    In this study we verified the method of impulse acoustic microscopy as a tool for scaffold evaluation in tissue engineering investigation. Cellulose diacetate (CDA) non-woven 3D scaffold was used as a model object. Scanning electron microscopy and optical microscopy were used as reference methods in order to realize feasibility of acoustic microscopy method in a regenerative medicine field. Direct comparison of the different methods was carried out.

  11. THEORETICAL AND METHODOLOGICAL APPROACHES TO REGIONAL COMPETITION INVESTIGATION

    Directory of Open Access Journals (Sweden)

    A.I. Tatarkin

    2006-03-01

    Full Text Available The article is dedicated to theoretical-methodological issues of regional economy competitiveness investigation. Economic essence of regional competitiveness is analyzed, its definition is given. The factors that determine relations of competition on medium and macrolevels are proved. The basic differences between world-economical and inter-regional communications are formulated. The specific features of globalization processes as form of competitive struggle are considered.

  12. Investigation of anticancer potential of hypophyllanthin and phyllanthin against breast cancer by in vitro and in vivo methods

    Directory of Open Access Journals (Sweden)

    Madhukiran Parvathaneni

    2014-02-01

    Full Text Available Objective: To investigate the in vitro and in vivo anticancer activities of hypophyllanthin and phyllanthin isolated from Phyllanthus amarus Schum & Thonn against breast cancer. Methods: In vitro anticancer activity was evaluated against two cell lines (MCF-7 and MDAMB-231 using MTT assay. In vivo anticancer activity was tested using Sprague-Dawley rats with N-methyl-N-nitrosourea induced mammary cancer. Results: In vitro studies demonstrated a dose-dependent inhibitory effect on cell growth with IC50 values of (35.18依1.48 µg/mL (hypophyllanthin and (32.51依0.95 µg/mL (phyllanthin for MCF-7; (38.74 依1.24 (hypophyllanthin and (32.2依1.17 (phyllanthin for MDA-MB-231 breast cancer cell lines. Tumor weights per group at doses of 5 and 10 mg/kg/day for hypophyllanthin (12.82 and 12.06 g and phyllanthin (11.95 and 8.87 g treated groups were significantly (P<0.001 lower than untreated N-methyl-N-nitrosourea group (35.85. Conclusions: Results of the present research work indicated that the isolated lignan compounds, hypophyllanthin and phyllanthin showed significant anticancer activities against breast cancer, in vitro and in vivo.

  13. Investigation into Effects of Scanning Speed on in Vitro Biocompatibility of Selective Laser Melted 316L Stainless Steel Parts

    Directory of Open Access Journals (Sweden)

    Shang Yitong

    2017-01-01

    Full Text Available In recent years, selective laser melting (SLM has gained an important place in fabrication due to their strong individualization which cannot be manufactured using conventional processes such as casting or forging. By proper control of the SLM processing parameters, characteristics of the alloy can be optimized. In the present work, 316L stainless steel (SS, as a widely used biomedical material, is investigated in terms of the effects of scanning speed on in vitro biocompatibility during SLM process. Cytotoxicity assay is adopted to assess the in vitro biocompatibility. The results show the scanning speed strongly affects the in vitro biocompatibility of 316L SS parts and with prolongs of incubation time, the cytotoxicity increase and the in vitro biocompatibility gets worse. The optimal parameters are determined as follows: scanning speed of 900 mm/s, laser power of 195 W, hatch spacing of 0.09 mm and layer thickness of 0.02 mm. The processing parameters lead to the change of surface morphology and microstructures of samples, which can affect the amount of toxic ions release, such as Cr, Mo and Co, that can increase risks to patient health and reduce the biocompatibility.

  14. An in vitro--in silico--in vivo approach in biopharmaceutical drug characterization: metformin hydrochloride IR tablets.

    Science.gov (United States)

    Beloica, S; Cvijić, S; Homšek, I; Bogataj, M; Parojčić, J

    2015-07-01

    The integrated in vitro--in silico--in vivo approach has emerged into a biopharmaceutical toolkit that could accelerate drug development and improve drug product clinical performance in patients. In the present study, the influence of physiologically based media and dynamic dissolution testing on drug release from two metformin hydrochloride immediate release products with proven bioequivalence was tested. Metformin-specific physiologically based pharmacokinetic (PBPK) model was developed based on a range of literature or in silico predicted data using gastrointestinal simulation technology implemented in the Simcyp software package. Various approaches were employed in order to estimate the human effective permeability which was used as input for metformin plasma profile simulation. Influence of the rate and extent of metformin dissolution on drug absorption was evaluated. Both convolution and deconvolution approaches were used in order to establish a correlation between the in vitro and in vivo data. The results obtained indicate that physiologically based dissolution media and glass bead dissolution device exhibit certain advantages over the compendial dissolution apparatus and simple buffers which tended to be over-discriminative. Gastrointestinal simulation technology implemented in the Simcyp Simulator was successfully used in developing drug-specific PBPK model for metformin. Simulations indicate that in vitro dissolution kinetics has no significant effect on metformin absorption, if more than 65% of drug is released in 1 hour. Level A in vitro-in vivo correlation was obtained using both convolution and deconvolution approaches.

  15. AnIn-vitroInvestigation of Swelling Controlled Delivery of Insulin from Egg Albumin Nanocarriers.

    Science.gov (United States)

    Mahobia, Swati; Bajpai, Jaya; Bajpai, Anil Kumar

    2016-01-01

    The aim of the present work was to prepare and characterize biopolymer nanocarriers and evaluate their suitability in possible oral delivery of insulin. The egg albumin biopolymer was used to prepare nanoparticles which were further characterized by Fourier transformed Infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), zeta potential, Dynamic Light scattering (DLS) and cytotoxicity. From the characterization studies the size of the nanoparticles washemoly found to lie in the range 20-80 nm with surface charge of -23 mV and also offering extremely fair biocompatibility.. The in-vitro biocompatibility of the prepared nanocarriers was judged by BSA adsorption test and haemolysis assay. The in vitro release kinetics of the insulin loaded nanoparticles was studied in phosphate buffer saline (PBS) solution, and the influence of various factors such as pH, temperature and simulated physiological fluids was studied on the controlled release of insulin.

  16. Novel approach to investigation of semiconductor MOCVD by microreactor technology

    Science.gov (United States)

    Konakov, S. A.; Krzhizhanovskaya, V. V.

    2017-11-01

    Metal-Organic Chemical Vapour Deposition is a very complex technology that requires further investigation and optimization. We propose to apply microreactors to (1) replace multiple expensive time-consuming macroscale experiments by just one microreactor deposition with many points on one substrate; (2) to derive chemical reaction rates from individual deposition profiles using theoretical analytical solution. In this paper we also present the analytical solution of a simplified equation describing the deposition rate dependency on temperature. It allows to solve an inverse problem and to obtain detailed information about chemical reaction mechanism of MOCVD process.

  17. Preparation and In vitro Investigation of Chitosan Compressed Tablets for Colon Targeting

    Directory of Open Access Journals (Sweden)

    Negar Bashardoust

    2011-12-01

    Full Text Available Purpose: The aim of the present study was minimizing the drug release in upper gastro intestinal tract and targeting to colon by using the principles of compression coat. Methods: Compression coated tablets of Ibuprofen were prepared by direct compression method using chitosan (300, 250, 200 & 175 mg. Tablets were evaluated for their physicochemical properties and in vitro drug release studies. In vitro drug release studies were performed with and without rat caecal contents. Results: In the rat caecal contents tablets showed enhanced drug release due to degradation of chitosan coat by colonic colonic enzymes. The in vitro release studies in pH-6.8 phosphate buffer containing 2% w/v of rat caecal contents showed the cumulative percentage release of Ibuprofen after 26h as 31.94% ±0.59, 67.89% ± 0.45 and 55.87 % ± 0.45 and 82.52 % ± 0.92 respectively. Coatthickness and amount of chitosan controls the release rate. Formulations are best fitted with Korsmeyer-Peppas kinetics and mechanism of drug release was non-Fickian. FTIR studies reveals there is no drug-polysaccharide interaction. F1 formulation was a promising system for drug targeting to colon. Conclusion: Based on the obtained results chitosan as a press coat could target ibuprofen to the colon.

  18. A new chemical approach to optimize the in vitro SPF method on the HD6 PMMA plate.

    Science.gov (United States)

    Marguerie, S; Pissavini, M; Baud, A; Carayol, T; Doucet, O

    2012-01-01

    In a previous study, we demonstrated that control of the roughness of molded PMMA plates improves in vitro SPF reproducibility. However, in vitro/vivo deviations are still observed. Sunscreens show different behavior during spreading on the HD6 surface according to the formulation, resulting in a more or less homogenous distribution. The hydrophilic nature of HD6 appears to contribute significantly during spreading. Two different sunscreens offering a homogenous and non-homogenous distribution were investigated to check if the interfacial tension between product and substrate has a real influence on the spreading quality. Using microscopic observations, we attempted to correlate the in vitro SPF results with the product's spreading property. In order to reduce this interfacial tension, an HD6 pretreatment with an amphoteric surfactant, cocamidopropyl betain, was performed. In vitro SPF on "pretreated HD6" was examined using a cohort of 30 products. This pretreatment led to reliable results, demonstrating good association with the in vivo SPF.

  19. A Research Strategy for Investigating Business Process Management Approaches

    Directory of Open Access Journals (Sweden)

    James Gibson

    2005-11-01

    Full Text Available We are witnessing a revolution in industry which, if successful, will change forever how business systems are developed and the type of staff required. This paradigm shift has only recently become possible as business process conceptual understanding evolved, technologies have matured and higher abstraction levels have become possible. Industry leads Business Processing Systems research as it has the strategic imperative and resources to be effective. Academic research is faced with three challenges: firstly, how to do effective research in an area of such broad scope, secondly, how to make research relevant to practice, thirdly how to spend limited resources effectively. This paper defines the research framework for effective academic research at the University of Wollongong by the Software Effective Process group. Effective research is enabled by co-ordinating research based on the primacy of the business model and its resultant effective representation in executable systems. The framework aims to build a core research team, promote strong synergy with existing research areas, and create academic and industry relevant research.. We report on the results to date of our pilot program and seek feedback and advice to help us refine our approach. A major Australian project is utilising a new software development lifecycle for ‘system of systems’ development which has arisen out of this research strategy. Later papers will report on both the theoretical basis and practical impacts of this work and other research by the group.

  20. Investigating reasons for CPAP adherence in adolescents: a qualitative approach.

    Science.gov (United States)

    Prashad, Priya S; Marcus, Carole L; Maggs, Jill; Stettler, Nicolas; Cornaglia, Mary A; Costa, Priscilla; Puzino, Kristina; Xanthopoulos, Melissa; Bradford, Ruth; Barg, Frances K

    2013-12-15

    Adolescents with obstructive sleep apnea syndrome (OSAS) represent an important but understudied subgroup of long-term continuous positive airway pressure (CPAP) users. The purpose of this qualitative study was to identify factors related to adherence from the perspective of adolescents and their caregivers. Individual open-ended, semi-structured interviews were conducted with adolescents (n = 21) and caregivers (n = 20). Objective adherence data from the adolescents' CPAP machines during the previous month was obtained. Adolescents with different adherence levels and their caregivers were asked their views on CPAP. Using a modified grounded theory approach, we identified themes and developed theories that explained the adolescents' adherence patterns. Adolescent participants (n = 21) were aged 12-18 years, predominantly male (n = 15), African American (n = 16), users of CPAP for at least one month. Caregivers were mainly mothers (n = 17). Seven adolescents had high use (mean use 381 ± 80 min per night), 7 had low use (mean use 30 ± 24 min per night), and 7 had no use during the month prior to being interviewed. Degree of structure in the home, social reactions, mode of communication among family members, and perception of benefits were issues that played a role in CPAP adherence. Understanding the adolescent and family experience of using CPAP may be key to increasing adolescent CPAP adherence. As a result of our findings, we speculate that health education, peer support groups, and developmentally appropriate individualized support strategies may be important in promoting adherence. Future studies should examine these theories of CPAP adherence.

  1. New experimental approaches to investigate the fission dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Benlliure, J., E-mail: j.benlliure@usc.es; Rodríguez-Sánchez, J. L.; Alvarez-Pol, H.; Ayyad, Y.; Cortina-Gil, D.; Paradela, C.; Pietras, B.; Ramos, D.; Vargas, J. [Universidade de Santiago de Compostela, 15782 Santiago de Compostela (Spain); Audouin, L.; Boutoux, G. [Institut de Physique Nucléaire d’Orsay, F-91406 Orsay (France); Bélier, G.; Chatillon, A.; Gorbinet, T.; Laurent, B.; Martin, J.-F.; Pellereau, E.; Taïeb, J. [CEA, DAM, DIF, F-91297 Arpajon (France); Casarejos, E. [Universidad de Vigo, E-36200 Vigo (Spain); Heinz, A. [Chalmers University of Technology, SE-412 96 Gothenburg (Sweden); and others

    2016-07-07

    The first ever achieved full identification of both fission fragments, in atomic and mass number, made it possible to define new observables sensitive to the fission dynamics along the fission path up to the scission point. Moreover, proton-induced fission of {sup 208}Pb at high energies offers optimal conditions for the investigation of dissipative, and transient effects, because of the high-excitation energy of the fissioning nuclei, its low angular momentum, and limited shape distortion by the reaction. In this work we show that the charge distribution of the final fission fragments can constrain the ground-to-saddle dynamics while the mass distribution is sensitive to the dynamics until the scission point.

  2. New Modeling Approaches to Investigate Cell Signaling in Radiation Response

    Science.gov (United States)

    Plante, Ianik; Cucinotta, Francis A.; Ponomarev, Artem L.

    2011-01-01

    Ionizing radiation damages individual cells and tissues leading to harmful biological effects. Among many radiation-induced lesions, DNA double-strand breaks (DSB) are considered the key precursors of most early and late effects [1] leading to direct mutation or aberrant signal transduction processes. In response to damage, a flow of information is communicated to cells not directly hit by the radiation through signal transduction pathways [2]. Non-targeted effects (NTE), which includes bystander effects and genomic instability in the progeny of irradiated cells and tissues, may be particularly important for space radiation risk assessment [1], because astronauts are exposed to a low fluence of heavy ions and only a small fraction of cells are traversed by an ion. NTE may also have important consequences clinical radiotherapy [3]. In the recent years, new simulation tools and modeling approaches have become available to study the tissue response to radiation. The simulation of signal transduction pathways require many elements such as detailed track structure calculations, a tissue or cell culture model, knowledge of biochemical pathways and Brownian Dynamics (BD) propagators of the signaling molecules in their micro-environment. Recently, the Monte-Carlo simulation code of radiation track structure RITRACKS was used for micro and nano-dosimetry calculations [4]. RITRACKS will be used to calculate the fraction of cells traversed by an ion and delta-rays and the energy deposited in cells in a tissue model. RITRACKS also simulates the formation of chemical species by the radiolysis of water [5], notably the .OH radical. This molecule is implicated in DNA damage and in the activation of the transforming growth factor beta (TGF), a signaling molecule involved in NTE. BD algorithms for a particle near a membrane comprising receptors were also developed and will be used to simulate trajectories of signaling molecules in the micro-environment and characterize autocrine

  3. Biotransformation of the mycotoxin enniatin B1 in pigs: A comparative in vitro and in vivo approach.

    Science.gov (United States)

    Ivanova, Lada; Uhlig, Silvio; Devreese, Mathias; Croubels, Siska; Fæste, Christiane Kruse

    2017-07-01

    Enniatins (ENNs) are hexadepsipeptidic mycotoxins produced by Fusarium species. They occur in mg/kg levels in grain from Northern climate areas. Major ENNs such as ENN B and B1 have shown considerable cytotoxicity in different in vitro test systems. To adequately assess exposure and in vivo toxicity the toxicokinetic properties need to be investigated. The present study describes the metabolism of ENN B1 both in vitro and in vivo in pigs, comparing metabolites found in vitro in experiments with liver microsomes from different pig strains to those found in the plasma of pigs after single oral or intravenous application of ENN B1. Metabolites of hepatic biotransformation were tentatively identified and characterised by high performance liquid chromatography coupled to ion trap and high-resolution mass spectrometry, as well as chemical derivatisations. Kinetic parameters of metabolite formation and elimination were determined. Metabolite formation was higher when ENN B1 was absorbed from the gut compared to intravenous administration indicating pre-systemic metabolism of ENN B1 after oral uptake. The in vitro approach resulted in the detection of ten ENN B1 metabolites, while six were detected in in vivo samples. The putative ENN B1 metabolites were products of hydroxylation, carbonylation, carboxylation and oxidative demethylation reactions. Copyright © 2017. Published by Elsevier Ltd.

  4. In Vitro and Ex Vivo Approaches to Evaluate Next-Generation Tobacco and Non-Tobacco Products on Human Blood Platelets.

    Science.gov (United States)

    Spinelli, Sherry L; Lannan, Katie L; Loelius, Shannon G; Phipps, Richard P

    2017-03-01

    Human blood platelets are major hemostatic regulators in the circulation and important in the mediation of chronic inflammation and immunomodulation. They are key elements that promote cardiovascular pathogenesis that leads to atherosclerosis, thrombosis, myocardial infarction, and stroke. New information on tobacco use and platelet dysregulation shows that these highly understudied vascular cells are dysregulated by tobacco smoke. Thus, platelet function studies should be an important consideration for the evaluation of existing and next-generation tobacco and non-tobacco products. Novel in vitro approaches are being sought to investigate these products and their influence on platelet function. Platelets are ideally suited for product assessment, as robust and novel in vitro translational methods are available to assess platelet function. Furthermore, the use of human biological systems has the advantage that risk predictions will better reflect the human condition.

  5. Investigating international new product diffusion speed: A semiparametric approach

    KAUST Repository

    Hartman, Brian M.

    2012-06-01

    Global marketing managers are interested in understanding the speed of the new product diffusion process and how the speed has changed in our ever more technologically advanced and global marketplace. Understanding the process allows firms to forecast the expected rate of return on their new products and develop effective marketing strategies. The most recent major study on this topic [Marketing Science 21 (2002) 97-114] investigated new product diffusions in the United States.We expand upon that study in three important ways. (1) Van den Bulte notes that a similar study is needed in the international context, especially in developing countries. Our study covers four new product diffusions across 31 developed and developing nations from 1980-2004. Our sample accounts for about 80% of the global economic output and 60% of the global population, allowing us to examine more general phenomena. (2) His model contains the implicit assumption that the diffusion speed parameter is constant throughout the diffusion life cycle of a product. Recognizing the likely effects on the speed parameter of recent changes in the marketplace, we model the parameter as a semiparametric function, allowing it the flexibility to change over time. (3) We perform a variable selection to determine that the number of internet users and the consumer price index are strongly associated with the speed of diffusion. © Institute of Mathematical Statistics, 2012.

  6. SCREENING APPROACHES FOR METHANE MITIGATING POTENTIAL OF TANNIN-CONTAINING PLANTS UNDER IN VITRO RUMEN ENVIRONMENT

    Directory of Open Access Journals (Sweden)

    A. Jayanegara

    2014-10-01

    Full Text Available The aim of the present study was to conduct univariate, bivariate and multivariate (principalcomponent analysis, PCA approaches in the screening of tannin-containing plants from variouscollection sites for their CH4 mitigating properties. Plant samples were obtained from various collectionsites in different countries, i.e. Indonesia (n = 27 species, Mongolia (n = 14, Switzerland (n = 16 andGermany (n = 3. The plants were incubated in vitro with buffered-rumen fluid at 39oC for 24 h. Totalgas production was recorded as an indicator of feed quality and emission of CH4 was measured. Resultsshowed that, based on bivariate screening, generally, plants possessed low CH4 production had lowquality or low total gas production except Rhus typhina, i.e. 43 ml/200 mg DM. The loading plot of PCAshowed that all phenolic fractions were in the opposite direction with CH4 and total gas production.Plants clustered together in reverse direction to that of CH4 were Bergenia crassifolia root and leaf,Swietenia mahagoni, Clidemia hirta, Peltiphyllum peltatum, Acacia villosa and R. typhina. It wasconluded that, for tannin-containing plants, screenings based on univariate, bivariate and multivariateapproaches in relation to ruminal CH4 emission led to similar results.

  7. Isolation of hydroxytyrosol from olive leaves extract, radioiodination and investigation of bioaffinity using in vivo/in vitro methods

    Energy Technology Data Exchange (ETDEWEB)

    Ozkan, M.; Biber Muftuler, F.Z.; Kilcar, A. Yurt; Medine, E.I.; Unak, P. [Ege Univ., Izmir (Turkey). Dept. of Nuclear Applications

    2013-11-01

    It is known that medicinal plants like olive have biological activities due to their flavonoid content such as olueropein, tyrosol, hydroxytyrosol etc. In current study, hydroxytrosol (HT) which is one of the major phenolic compounds in olive, olive leaves and olive oil, was isolated after methanol extraction and purification of olive leaves which are grown in the northern Anatolia region of Turkey. The isolated HT was radiolabeled with {sup 131}I ({sup 131}I-HT) and the bioaffinity of this radiolabeled component of olive leaves extract was investigated by using in vivo/in vitro methods. It was found that HT could be radiolabeled with {sup 131}I in yields of 95.6 {+-} 4.4% (n = 8), and in vivo studies showed that {sup 131}I-HT is taken up by urinary bladder, stomach, small intestine, large intestine, breast and prostate. Significant incorporation of activity was observed in cell lines via in vitro studies. (orig.)

  8. The development and characterization of a human mesothelioma in vitro 3D model to investigate immunotoxin therapy.

    Directory of Open Access Journals (Sweden)

    Xinran Xiang

    2011-01-01

    Full Text Available Tumor microenvironments present significant barriers to penetration by antibodies and immunoconjugates. Tumor microenvironments, however, are difficult to study in vitro. Cells cultured as monolayers exhibit less resistance to therapy than those grown in vivo and an alternative research model more representative of the in vivo tumor is more desirable. SS1P is an immunotoxin composed of the Fv portion of a mesothelin-specific antibody fused to a bacterial toxin that is presently undergoing clinical trials in mesothelioma.Here, we examined how the tumor microenvironment affects the penetration and killing activity of SS1P in a new three-dimensional (3D spheroid model cultured in vitro using the human mesothelioma cell line (NCI-H226 and two primary cell lines isolated from the ascites of malignant mesothelioma patients. Mesothelioma cells grown as monolayers or as spheroids expressed comparable levels of mesothelin; however, spheroids were at least 100 times less affected by SS1P. To understand this disparity in cytotoxicity, we made fluorescence-labeled SS1P molecules and used confocal microscopy to examine the time course of SS1P penetration within spheroids. The penetration was limited after 4 hours. Interestingly, we found a significant increase in the number of tight junctions in the core area of spheroids by electron microscopy. Expression of E-Cadherin, a protein involved in the assembly and sealing of tight junctions and highly expressed in malignant mesothelioma, was found significantly increased in spheroids as compared to monolayers. Moreover, we found that siRNA silencing and antibody inhibition targeting E-Cadherin could enhance SS1P immunotoxin therapy in vitro.This work is one of the first to investigate immunotoxins in 3D tumor spheroids in vitro. This initial description of an in vitro tumor model may offer a simple and more representative model of in vivo tumors and will allow for further investigations of the microenvironmental

  9. Cochlear implants: in vitro investigation of electromagnetic interference at MR imaging--compatibility and safety aspects.

    Science.gov (United States)

    Teissl, C; Kremser, C; Hochmair, E S; Hochmair-Desoyer, I J

    1998-09-01

    In vitro experiments were performed to determine the compatibility of a cochlear implant at 0.2- and 1.5-T magnetic resonance (MR) imaging. Except for the torque at 1.5 T, all electromagnetic interferences remained within acceptable limits. MR imaging should be performed only if there is a strong medical indication, by following appropriate safety procedures. MR imaging at 0.2 T should be safe; at 1.5 T, however, the relative risks will have to be assessed.

  10. In vitro Culture of Naïve Human Bone Marrow Mesenchymal Stem Cells: A Stemness Based Approach

    Science.gov (United States)

    Pal, Bidisha; Das, Bikul

    2017-01-01

    Human bone marrow derived mesenchymal stem cells (BM-MSCs) resides in their niches in close proximity to hematopoietic stem cells (HSCs). These naïve MSCs have tremendous potential in regenerative therapeutics, and may also be exploited by cancer and infectious disease agents. Hence, it is important to study the physiological and pathological roles of naïve MSC. However, our knowledge of naïve MSCs is limited by lack of appropriate isolation and in vitro culture methods. Established culture methods use serum rich media, and serial passaging for retrospective isolation of MSCs. These primed MSCs may not reflect the true physiological and pathological roles of naive MSCs (Figure 1). Therefore, there is a strong need for direct isolation and in vitro culture of naïve MSCs to study their stemness (self-renewal and undifferentiated state) and developmental ontogeny. We have taken a niche-based approach on stemness to better maintain naïve MSCs in vitro. In this approach, stemness is broadly divided as niche dependent (extrinsic), niche independent (intrinsic) and niche modulatory (altruistic or competitive). Using this approach, we were able to maintain naïve CD271+/CD133+ BM-MSCs for 2 weeks. Furthermore, this in vitro culture system helped us to identify naïve MSCs as a protective niche site for Mycobacterium tuberculosis, the causative organism of pulmonary tuberculosis. In this review, we discuss the in vitro culture of primed vs. naïve human BM derived MSCs with a special focus on how a stemness based approach could facilitate the study of naïve BM-MSCs. PMID:28884113

  11. In vitro Culture of Naïve Human Bone Marrow Mesenchymal Stem Cells: A Stemness Based Approach.

    Science.gov (United States)

    Pal, Bidisha; Das, Bikul

    2017-01-01

    Human bone marrow derived mesenchymal stem cells (BM-MSCs) resides in their niches in close proximity to hematopoietic stem cells (HSCs). These naïve MSCs have tremendous potential in regenerative therapeutics, and may also be exploited by cancer and infectious disease agents. Hence, it is important to study the physiological and pathological roles of naïve MSC. However, our knowledge of naïve MSCs is limited by lack of appropriate isolation and in vitro culture methods. Established culture methods use serum rich media, and serial passaging for retrospective isolation of MSCs. These primed MSCs may not reflect the true physiological and pathological roles of naive MSCs (Figure 1). Therefore, there is a strong need for direct isolation and in vitro culture of naïve MSCs to study their stemness (self-renewal and undifferentiated state) and developmental ontogeny. We have taken a niche-based approach on stemness to better maintain naïve MSCs in vitro. In this approach, stemness is broadly divided as niche dependent (extrinsic), niche independent (intrinsic) and niche modulatory (altruistic or competitive). Using this approach, we were able to maintain naïve CD271+/CD133+ BM-MSCs for 2 weeks. Furthermore, this in vitro culture system helped us to identify naïve MSCs as a protective niche site for Mycobacterium tuberculosis, the causative organism of pulmonary tuberculosis. In this review, we discuss the in vitro culture of primed vs. naïve human BM derived MSCs with a special focus on how a stemness based approach could facilitate the study of naïve BM-MSCs.

  12. A multivariate approach linking reported side effects of clinical antidepressant and antipsychotic trials to in vitro binding affinities.

    Science.gov (United States)

    Michl, Johanna; Scharinger, Christian; Zauner, Miriam; Kasper, Siegfried; Freissmuth, Michael; Sitte, Harald H; Ecker, Gerhard F; Pezawas, Lukas

    2014-09-01

    The vast majority of approved antidepressants and antipsychotics exhibit a complex pharmacology. The mechanistic understanding of how these psychotropic medications are related to adverse drug reactions (ADRs) is crucial for the development of novel drug candidates and patient adherence. This study aims to associate in vitro assessed binding affinity profiles (39 compounds, 24 molecular drug targets) and ADRs (n=22) reported in clinical trials of antidepressants and antipsychotics (n>59.000 patients) by the use of robust multivariate statistics. Orthogonal projection to latent structures (O-PLS) regression models with reasonable predictability were found for several frequent ADRs such as nausea, diarrhea, hypotension, dizziness, headache, insomnia, sedation, sleepiness, increased sweating, and weight gain. Results of the present study support many well-known pharmacological principles such as the association of hypotension and dizziness with α1-receptor or sedation with H1-receptor antagonism. Moreover, the analyses revealed novel or hardly investigated mechanisms for common ADRs including the potential involvement of 5-HT6-antagonism in weight gain, muscarinic receptor antagonism in dizziness, or 5-HT7-antagonism in sedation. To summarize, the presented study underlines the feasibility and value of a multivariate data mining approach in psychopharmacological development of antidepressants and antipsychotics. Copyright © 2014. Published by Elsevier B.V.

  13. A structure-based Multiple-Instance Learning approach to predicting in vitro transcription factor-DNA interaction.

    Science.gov (United States)

    Gao, Zhen; Ruan, Jianhua

    2015-01-01

    Understanding the mechanism of transcriptional regulation remains an inspiring stage of molecular biology. Recently, in vitro protein-binding microarray experiments have greatly improved the understanding of transcription factor-DNA interaction. We present a method - MIL3D - which predicts in vitro transcription factor binding by multiple-instance learning with structural properties of DNA. Evaluation on in vitro data of twenty mouse transcription factors shows that our method outperforms a method based on simple-instance learning with DNA structural properties, and the widely used k-mer counting method, for nineteen out of twenty of the transcription factors. Our analysis showed that the MIL3D approach can utilize subtle structural similarities when a strong sequence consensus is not available. Combining multiple-instance learning and structural properties of DNA has promising potential for studying biological regulatory networks.

  14. Development of an integrated approach for comparison of in vitro and in vivo responses to particulate matter.

    Science.gov (United States)

    Breznan, Dalibor; Karthikeyan, Subramanian; Phaneuf, Marcelle; Kumarathasan, Prem; Cakmak, Sabit; Denison, Michael S; Brook, Jeffrey R; Vincent, Renaud

    2016-08-12

    ) approached the threshold for significance (p = 0.052) in a correlation within in vitro and in vivo models, with a ranking of fine particle (DWR1), minerals (TiO2, CRI) and coarse particles (SRM-, EHC-type) from low to high potency. Integration of in vitro endpoints shows promise in determining adverse outcomes of particle exposures in vivo. The devised data reduction and computational approach will prove useful in the development of models for assessment of hazard potential of particles; however, distinct models may be needed for particles of different type, such as urban particles vs. mineral particles, nanomaterials.

  15. Salbutamol sulphate-ethylcellulose microparticles: formulation and in-vitro evaluation with emphasis on mathematical approaches

    Directory of Open Access Journals (Sweden)

    G Murtaza

    2009-10-01

    Full Text Available "n "nBackground and the purpose of the study: This study reports the laboratory optimization for the preparation of salbutamol sulphate-ethylcellulose microparticles by a non-solvent addition coacervation technique through adjustment of the ratio of salbutamol sulphate to ethylcellulose. The variation of drug release between the microparticles and tabletted microparticles was also investigated. "nMethods: In vitro release profiles of developed microparticles and tabletted microparticles were studied using USP XXIV dissolution apparatus I and II, respectively, in 450 ml double distilled water at 50 rpm maintained at 37°C. "nResults: White microparticles with no definite shape having good entrapment efficiency (96.68 to 97.83% and production yield (97.48 ± 1.21 to 98.35 ± 1.08% were obtained. In this investigation, initial burst effect was observed in the drug release behavior. The rate of drug release from microparticles decreased as the concentration of polyisobutylene was increased from 6% to 12% during microencapsulation. The release pattern of tabletted microparticles was affected significantly (p < 0.05 by the addition of hydroxy propyl methyl cellulose (HPMC as excepient and insignificantly (p > 0.05 by the type of dissolution media and stirring speed. Tabletted microparticles showed good stability and reproducibility. Ethylcellulose was found to be compatible with salbutamol sulphate. The drug release from all formulations was best fit to Higuchi's equation and the mechanism of drug release was anomalous diffusion from all formulations. "nConclusion: The results of this study suggest that by using ethylcellulose it is possible to design a single-unit, sustained-release oral dosage form of salbutamol sulphate for indication of twice a day.

  16. Building a Tiered Approach to In Vitro Predictive Toxicity Screening: A Focus on Assays with In Vivo Relevance

    Science.gov (United States)

    McKim, James M

    2010-01-01

    One of the greatest challenges facing the pharmaceutical industry today is the failure of promising new drug candidates due to unanticipated adverse effects discovered during preclinical animal safety studies and clinical trials. Late stage attrition increases the time required to bring a new drug to market, inflates development costs, and represents a major source of inefficiency in the drug discovery/development process. It is generally recognized that early evaluation of new drug candidates is necessary to improve the process. Building in vitro data sets that can accurately predict adverse effects in vivo would allow compounds with high risk profiles to be deprioritized, while those that possess the requisite drug attributes and a lower risk profile are brought forward. In vitro cytotoxicity assays have been used for decades as a tool to understand hypotheses driven questions regarding mechanisms of toxicity. However, when used in a prospective manner, they have not been highly predictive of in vivo toxicity. Therefore, the issue may not be how to collect in vitro toxicity data, but rather how to translate in vitro toxicity data into meaningful in vivo effects. This review will focus on the development of an in vitro toxicity screening strategy that is based on a tiered approach to data collection combined with data interpretation. PMID:20053163

  17. Investigation on biological properties of tacrolimus-loaded poly(1,3-trimethylene carbonate) in vitro

    Science.gov (United States)

    Hou, Ruixia; Wu, Leigang; Wang, Jin; Huang, Nan

    2010-06-01

    The drug-eluting stents have been regarded as a milestone in inhibiting the restenosis of coronary arteries. However, adverse reactions caused by bare-metal stents and non-biodegradable polymer coatings may result in some clinical problems. In this study, a new tacrolimus-eluting stent coated with biodegradable poly(1,3-trimethylene carbonate) (PTMC) is developed. The structures are characterized by Fourier transform infrared (FTIR) analysis, and the wettability is measured by contact angle assay. The biological behaviors are evaluated by the in vitro platelets adhesion test, APTT test, the human umbilical cord artery smooth muscle cells (HUCASMCs), 4',6-diamidine-2-phenylindole (DAPI) and actin immunofluorescence staining, MTT colorimetric assay. These results show that after blending tacrolimus into PTMC, the anticoagulant behavior is improved, and the adhesion and proliferation of HUCASMCs on samples are inhibited. This work aims to find one kind of surface erosion biodegradable polymers that can be applied as drug-eluting stent coatings.

  18. Investigation of Gelatin Polypeptides of Jellyfish (Rhopilema esculentum for Their Antioxidant Activity in vitro

    Directory of Open Access Journals (Sweden)

    Yong-Liang Zhuang

    2010-01-01

    Full Text Available Jellyfish gelatin was hydrolyzed by different proteases to obtain antioxidative polypeptides. The gelatin hydrolysate obtained by progressive hydrolysis using trypsin and Properase E exhibited the highest hydrolysis degree and antioxidant activity. Three series of gelatin polypeptides (SCP1, SCP2 and SCP3 were obtained by ultrafiltrating the gelatin hydrolysate through molecular mass cut-off membranes of 10, 6 and 2 kDa, respectively. Amino acid composition analysis showed that SCP3 had the highest total hydrophobic amino acid content. The in vitro antioxidant tests demonstrated that SCP2 had the strongest hydroxyl radical and hydrogen peroxide scavenging activities and metal chelating ability, while SCP3 showed the highest reducing power, antioxidant activity in linoleic acid emulsion system and superoxide anion radical scavenging activity. The results support the feasibility of jellyfish gelatin as a natural antioxidant polypeptide provider, and enzymatic hydrolysis and ultrafiltration could be potent future processing technologies to utilize the abundant jellyfish resource.

  19. Reliability and validation of in vitro lumbar spine height measurements using musculoskeletal ultrasound: A preliminary investigation.

    Science.gov (United States)

    Sobczak, Stéphane; Dugailly, Pierre-Michel; Gilbert, Kerry K; Hooper, Troy L; Sizer, Phillip S; James, C Roger; Poortmans, Bernard; Matthijs, Omer C; Brismée, Jean-Michel

    2016-01-01

    Stadiometry measures total trunk height variations but cannot quantify individual spinal segment height changes. Different methods exist to measure both intervertebral disc and lumbar spine height (LSH) variations but they are either limited by radiation exposure or cost. Musculoskeletal ultrasound could be a valuable alternative to measure spinal segmental height changes as a result of intervention. To validate the use of musculoskeletal ultrasound (MSU) and new anatomical landmark references used in assessing inter-mammillary distances (IMD) and LSH changes resulting from lumbar spine traction. Two unembalmed cadaveric lumbar spines were extracted to assess (1) the reliability and validity of MSU, as compared to caliper, for measuring in vitro IMD and LSH using alternative anatomical landmarks than previously reported, and (2) the reliability of MSU for measuring in vitro IMD and lumbar spine height changes recorded during standardized mechanical traction up to 1.20 cm. Intra- and inter-rater reliability of musculoskeletal ultrasound for within and between sessions and for all experimental design, Standard Error ranged from 0.01 to 0.02 and from 0.03 to 0.04 cm for IMDs and LSHs, respectively. Root Mean Square Errors ranged from 1.6 to 6.8% and from 1 to 1.1% for IMDs and LSHs, respectively and mean ICC ranged from 0.98 to 1 for LSH. During traction, mean lumbar spine height measurement change using MSU was 1.15 ± 0.03 cm. Bland and Altman plots demonstrated confidence intervals included in the limits of agreement. Nevertheless, there were significant differences (plumbar spine height between caliper and ultrasound measurements. Musculoskeletal ultrasound overestimated distances of about 5.5 ± 1.5%. Musculoskeletal ultrasound is reliable and accurate for measuring intersegmental spinal distances and lumbar spine height with an apparent slight overestimation of distances. Based on mean differences, ultrasound technology seems to be valid for measuring lumbar

  20. Discovering probiotic microorganisms: in vitro, in vivo, genetic and omics approaches

    Directory of Open Access Journals (Sweden)

    Konstantinos ePapadimitriou

    2015-02-01

    Full Text Available Over the past decades the food industry has been revolutionized towards the production of functional foods due to an increasing awareness of the consumers on the positive role of food in wellbeing and health. By definition probiotic foods must contain live microorganisms in adequate amounts so as to be beneficial for the consumer’s health. There are numerous probiotic foods marketed today and many probiotic strains are commercially available. However, the question that arises is how to determine the real probiotic potential of microorganisms. This is becoming increasingly important, as even a superficial search of the relevant literature reveals that the number of proclaimed probiotics is growing fast. While the vast majority of probiotic microorganisms are food-related or commensal bacteria that are often regarded as safe, probiotics from other sources are increasingly being reported raising possible regulatory and safety issues. Potential probiotics are selected after in vitro or in vivo assays by evaluating simple traits such as resistance to the acidic conditions of the stomach or bile resistance, or by assessing their impact on complicated host functions such as immune development, metabolic function or gut-brain interaction. While final human clinical trials are considered mandatory for communicating health benefits, rather few strains with positive studies have been able to convince legal authorities with these health claims. Consequently, concern has been raised about the validity of the workflows currently used to characterize probiotics. In this review we will present an overview of the most common assays employed in screening for probiotics, highlighting the potential strengths and limitations of these approaches. Furthermore, we will focus on how the advent of omics technologies has reshaped our understanding of the biology of probiotics, allowing the exploration of novel routes for screening and studying such microorganisms.

  1. Physiological vortices in the sinuses of Valsalva: An in vitro approach for bio-prosthetic valves.

    Science.gov (United States)

    Toninato, Riccardo; Salmon, Jacob; Susin, Francesca Maria; Ducci, Andrea; Burriesci, Gaetano

    2016-09-06

    The physiological flow dynamics within the Valsalva sinuses, in terms of global and local parameters, are still not fully understood. This study attempts to identify the physiological conditions as closely as possible, and to give an explanation of the different and sometime contradictory results in literature. An in vitro approach was implemented for testing porcine bio-prosthetic valves operating within different aortic root configurations. All tests were performed on a pulse duplicator, under physiological pressure and flow conditions. The fluid dynamics established in the various cases were analysed by means of 2D Particle Image Velocimetry, and related with the achieved hydrodynamic performance. Each configuration is associated with substantially different flow dynamics, which significantly affects the valve performance. The configuration most closely replicating healthy native anatomy was characterised by the best hemodynamic performance, and any mismatch in size and position between the valve and the root produced substantial modification of the fluid dynamics downstream of the valve, hindering the hydrodynamic performance of the system. The worst conditions were observed for a configuration characterised by the total absence of the Valsalva sinuses. This study provides an explanation for the different vortical structures described in the literature downstream of bioprosthetic valves, enlightening the experimental complications in valve testing. Most importantly, the results clearly identify the fluid mechanisms promoted by the Valsalva sinuses to enhance the ejection and closing phases, and this study exposes the importance of an optimal integration of the valve and root, to operate as a single system. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. In vitro and in vivo investigation of taste-masking effectiveness of Eudragit E PO as drug particle coating agent in orally disintegrating tablets.

    Science.gov (United States)

    Drašković, Milica; Medarević, Djordje; Aleksić, Ivana; Parojčić, Jelena

    2017-05-01

    Considering that bitter taste of drugs incorporated in orally disintegrating tablets (ODTs) can be the main reason for avoiding drug therapy, it is of the utmost importance to achieve successful taste-masking. The evaluation of taste-masking effectiveness is still a major challenge. The objective of this study was to mask bitter taste of the selected model drugs by drug particle coating with Eudragit ® E PO, as well as to evaluate taste-masking effectiveness of prepared ODTs using compendial dissolution testing, dissolution in the small-volume shake-flask assembly and trained human taste panel. Model drugs were coated in fluidized bed. Disintequik™ ODT was used as a novel co-processed excipient for ODT preparation. Selected formulations were investigated in vitro and in vivo using techniques for taste-masking assessment. Significantly slower drug dissolution was observed from tablets with coated drug particles during the first 3 min of investigation. Results of in vivo taste-masking assessment demonstrated significant improvement in drug bitterness suppression in formulations with coated drug. Strong correlation between the results of drug dissolution in the small-volume shake-flask assembly and in vivo evaluation data was established (R ≥ 0.970). Drug particle coating with Eudragit ® E PO can be a suitable approach for bitter taste-masking. Strong correlation between in vivo and in vitro results implicate that small-volume dissolution method may be used as surrogate for human panel taste-masking assessment, in the case of physical taste-masking approach application.

  3. The reconstructed skin model as a new tool for investigating in vitro dermal fillers: increased fibroblast activity by hyaluronic acid.

    Science.gov (United States)

    Girardeau-Hubert, Sarah; Teluob, Séverine; Pageon, Hervé; Asselineau, Daniel

    2015-01-01

    Clinical studies on dermal fillers have essentially focused upon visible improvement of skin quality and any eventual side effects, whereas very little is known about their detailed biological effects. New skin equivalent models were created to investigate the biological impact of hyaluronic acid (HA) fillers on the dermal compartment in vitro. Two different reconstructed skin models were developed to incorporate HA within the collagen fibers. In the mixed model, HA was distributed throughout the whole collagen gel whereas the HA was concentrated in the center of collagen gel in the inclusion model. A comparison of the addition of fillers in two models of reconstructed skin has permitted a better understanding of the biological impact of HA fillers. Protein profiling of supernatants from both models suggested a regulation of MMP-1 secretion by fibroblasts as a function of HA volume, distribution in the dermis and degree of cross-linking. Immunostaining of the inclusion model revealed increased production of type I and III procollagens close to the cross-linked HA. Fibroblasts located in this area showed a fusiform morphology as well as an increase in -smooth actin expression. The observed increase in collagen production may thus result in part from tension in fibroblasts surrounding the cross-linked HA. The inclusion reconstructed skin model, as compared to the mixed model, presented here, appears to be a useful tool for investigating the properties of various fillers in vitro and closer to the in vivo situation; our results show that HA fillers promote in vitro remodeling of the dermis by fibroblasts.

  4. Phytochemical and in vitro and in vivo biological investigation on the antihypertensive activity of mango leaves (Mangifera indica L.).

    Science.gov (United States)

    Ronchi, Silas Nascimento; Brasil, Girlandia Alexandre; do Nascimento, Andrews Marques; de Lima, Ewelyne Miranda; Scherer, Rodrigo; Costa, Helber B; Romão, Wanderson; Boëchat, Giovanna Assis Pereira; Lenz, Dominik; Fronza, Marcio; Bissoli, Nazaré Souza; Endringer, Denise Coutinho; de Andrade, Tadeu Uggere

    2015-10-01

    The aim of this study was to investigate the antihypertensive effect of leaves Mangifera indica L. using in vitro and in vivo assays. The ethanol extract of leaves of M. indica was fractionated to dichloromethanic, n-butyl alcohol and aqueous fractions. The chemical composition of ethanolic extract and dichloromethanic fraction were evaluated by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Antioxidant activity was evaluated in the DPPH scavenging activity assay. Angiotensin-converting enzyme (ACE) inhibitory activity was investigated using in vitro and in vivo assays. The chronic antihypertensive assay was performed in spontaneously hypertensive rats (SHRs) and Wistar rats treated with enalapril (10 mg/kg), dichloromethanic fraction (100 mg/kg; twice a day) or vehicle control for 30 days. The baroreflex sensitivity was evaluated through the use of sodium nitroprusside and phenylephrine. Cardiac hypertrophy was evaluated by morphometric analysis. The dichloromethanic fraction exhibited the highest flavonoid, total phenolic content and high antioxidant activity. Dichloromethanic fraction elicited ACE inhibitory activity in vitro (99 ± 8%) similar to captopril. LC-MS/MS analysis revealed the presence of ferulic acid (48.3 ± 0.04 µg/g) caffeic acid (159.8 ± 0.02 µg/g), gallic acid (142.5 ± 0.03 µg/g), apigenin (11.0 ± 0.01 µg/g) and quercetin (203.3 ± 0.05 µg/g). The chronic antihypertensive effects elicited by dichloromethanic fraction were similar to those of enalapril, and the baroreflex sensitivity was normalized in SHR. Plasma ACE activity and cardiac hypertrophy were comparable with animals treated with enalapril. Dichloromethanic fraction of M. indica presented an antihypertensive effect, most likely by ACE inhibition, with benefits in baroreflex sensitivity and cardiac hypertrophy. Altogether, the results of the present study suggest that the dichloromethanic fraction of M. indica leaves may have potential as a promoting

  5. In vitro approaches to evaluate placental drug transport by using differentiating JEG-3 human choriocarcinoma cells.

    Science.gov (United States)

    Ikeda, Kenji; Utoguchi, Naoki; Tsutsui, Hidenobu; Yamaue, Satoko; Homemoto, Manami; Nakao, Erina; Hukunaga, Yumi; Yamasaki, Kyohei; Myotoku, Michiaki; Hirotani, Yoshihiko

    2011-02-01

    Human choriocarcinoma cells have been used as models for studying transcellular drug transport through placental trophoblasts. However, these models allow the transport of low-molecular-weight drugs through intercellular gap junctions. This study aimed at investigating the differentiation patterns of JEG-3 choriocarcinoma cells under different culture conditions and establishing the appropriate model of in vitro syncytiotrophoblast drug transport. Paracellular permeability was estimated by measuring the transepithelial electrical resistance (TEER) across JEG-3 cell layers. The mRNA expression levels of non-expressed in choriocarcinoma clone 1 (NECC1) and breast cancer resistance protein (BCRP), and those of E-cadherin (ECAD) and cadherin-11 (CDH11), which are adherens junction-associated proteins related to fusogenic ability of syncytiotrophoblasts differentiated from cytotrophoblasts, protein expression levels were considered as the differentiation signals. The highest TEER values were obtained in the JEG-3 cells cultured in the Dulbecco's modified Eagle's medium (DMEM)/Ham's F-12 (1:1) mixed medium (CS-C(®) ; Dainippon Sumitomo Pharma Co. Ltd., Osaka, Japan). By comparing the TEER values and the differentiation signals, the authors identified at least five JEG-3 cell-differentiation patterns. The differentiation pattern of JEG-3 cultured in CS-C resembled the syncytiotrophoblast-like differentiation signal characterizations in vivo. In conclusion, the syncytiotrophoblast-like models of differentiating JEG-3 cells cultured in CS-C might be appropriate for evaluating drug transport across the placental trophoblast. © 2010 The Authors. Basic & Clinical Pharmacology & Toxicology © 2010 Nordic Pharmacological Society.

  6. Investigation on in vitro dissolution rate enhancement of indomethacin by using a novel carrier sucrose fatty acid ester

    Directory of Open Access Journals (Sweden)

    Murthy Kolapalli Venkata

    2012-07-01

    Full Text Available Abstract Background and the purpose of the study The purpose of the present investigation was to characterize and evaluate solid dispersions (SD of indomethacin by using a novel carrier sucrose fatty acid ester (SFE 1815 to increase its in vitro drug release and further formulating as a tablet. Methods Indomethacin loaded SD were prepared by solvent evaporation and melt granulation technique using SFE 1815 as carrier in 1:0.25, 1:0.5 1:0.75 and 1:1 ratios of drug and carrier. Prepared SD and tablets were subjected to in vitro dissolution studies in 900 mL of pH 7.2 phosphate buffer using apparatus I at 100 rpm. The promising SD were further formulated as tablets using suitable diluent (DCL 21, Avicel PH 102 and pregelatinised starch to attain the drug release similar to that of SD.. The obtained dissolution data was subjected to kinetic study by fitting the data into various model independent models like zero order, first order, Higuchi, Hixon-Crowell and Peppas equations. Drug and excipient compatibility studies were confirmed by fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimetry and scanning electron microscopy. Results The in vitro dissolution data exhibited superior release from formulation S6 with 1:0.5 drug and carrier ratio using solvent evaporation technique than other SDs prepared at different ratio using solvent evaporation and melt granulation technique. The in vitro drug release was also superior to that of the physical mixtures prepared at same ratio and also superior to SD prepared using common carriers like polyvinyl pyrollidone and PEG 4000 by solvent evaporation technique. Tablets (T8 prepared with DCL21 as diluent exhibited superior release than the other tablets. The tablet formulation (T8 followed first order release with Non-Fickian release. Conclusion SFE 1815 a novel third generation carrier can be used for the preparation of SD for the enhancement of in vitro drug release of

  7. Magnetic core/shell nanoparticle thin films deposited by MAPLE: Investigation by chemical, morphological and in vitro biological assays

    Science.gov (United States)

    Cristescu, R.; Popescu, C.; Socol, G.; Iordache, I.; Mihailescu, I. N.; Mihaiescu, D. E.; Grumezescu, A. M.; Balan, A.; Stamatin, I.; Chifiriuc, C.; Bleotu, C.; Saviuc, C.; Popa, M.; Chrisey, D. B.

    2012-09-01

    We report on thin film deposition of nanostructured Fe3O4/oleic acid/ceftriaxone and Fe3O4/oleic acid/cefepime nanoparticles (core/shell/adsorption-shell) were fabricated by matrix assisted pulsed laser evaporation (MAPLE) onto inert substrates. The thin films were characterized by profilometry, Fourier transform infrared spectroscopy, atomic force microscopy, and investigated by in vitro biological assays. The biological properties tested included the investigation of the microbial viability and the microbial adherence to the glass coverslip nanoparticle film, using Gram-negative and Gram-positive bacterial strains with known antibiotic susceptibility behavior, the microbial adherence to the HeLa cells monolayer grown on the nanoparticle pellicle, and the cytotoxicity on eukaryotic cells. The proposed system, based on MAPLE, could be used for the development of novel anti-microbial materials or strategies for fighting pathogenic biofilms frequently implicated in the etiology of biofilm associated chronic infections.

  8. Investigation of the bystander effect in MRC5 cells after acute and fractionated irradiation in vitro

    OpenAIRE

    Shokouhozaman Soleymanifard; Mohammad Taghi Bahreyni Toossi; Roghayeh Kamran Samani; Shokoufeh Mohebbi

    2014-01-01

    Radiation-induced bystander effect (RIBE) has been defined as radiation responses observed in nonirradiated cells. It has been the focus of investigators worldwide due to the deleterious effects it induces in nonirradiated cells. The present study was performed to investigate whether acute or fractionated irradiation will evoke a differential bystander response in MRC5 cells. A normal human cell line (MRC5), and a human lung tumor cell line (QU-DB) were exposed to 0, 1, 2, and 4Gy of single a...

  9. Infant formula as a substrate for bifidogenesis: An in vitro investigation

    African Journals Online (AJOL)

    Pancreatin digestion preceded inoculation of the bifidobacteria biomass. The reference substrate was mature breast milk. This investigation showed important differences in bifidogenic effect between individual infant formulas and mature breast milk. In relation to dry biomass from the mature breast milk, dry biomass ...

  10. A comprehensive approach to in vitro functional evaluation of Ag/alginate nanocomposite hydrogels.

    Science.gov (United States)

    Stojkovska, Jasmina; Kostić, Danijela; Jovanović, Željka; Vukašinović-Sekulić, Maja; Mišković-Stanković, Vesna; Obradović, Bojana

    2014-10-13

    In this work, we present a comprehensive approach to evaluation of alginate microbeads with included silver nanoparticles (AgNPs) at the concentration range of 0.3-5mM for potential biomedical use by combining cytotoxicity, antibacterial activity, and silver release studies. The microbeads were investigated regarding drying and rehydration showing retention of ∼ 80-85% of the initial nanoparticles as determined by UV-vis and SEM analyses. Both wet and dry microbeads were shown to release AgNPs and/or ions inducing similar growth delays of Staphylococcus aureus and Escherichia coli at the total released silver concentrations of ∼ 10 μg/ml. On the other hand, these concentrations were highly toxic for bovine chondrocytes in conventional monolayer cultures while nontoxic when cultured in alginate microbeads under biomimetic conditions in 3D perfusion bioreactors. The applied approach outlined directions for further optimization studies demonstrating Ag/alginate microbeads as potentially attractive components of soft tissue implants as well as antimicrobial wound dressings. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Functional in vitro tension measurements of fascial tissue – a novel modified superfusion approach

    Science.gov (United States)

    Schleip, R.; Klingler, W.; Wearing, S.; Naylor, I.; Zuegel, M.; Hoppe, K.

    2016-01-01

    Introduction: While two laboratory techniques are commonly used to assess the tensile properties of muscle tissue, emerging evidence suggests that the fascial components of these tissues also serve an active role in force generation. Hence, we investigated whether these techniques are sensitive for assessment of fascial micromechanics. Methods: Force measurements on dissected fascial tissue were performed either using the classical immersion organ bath or using an improved superfusion approach simulating pulsed pharmacological triggers. Rat deep dorsal fascial strips as well as rat testicular capsule were pharmacologically challenged either with mepyramine or oxytocin. Results: The classical immersion technique yielded a lower force response to mepyramine than the superfusion method (median: 367.4 vs. 555.4µN/mm2). Pause in irrigation before application reduced irregularities during bolus application. The superfusion approach was improved further by the following points: The high sensitivity of the superfusion method to bolus addition was voided by deviation of fluid supply during bolus addition. Conclusion: Although both methods demonstrated pharmacologically induced contractile responses in lumbar fascia samples, the modified superfusion method may improve force registrations of slow contracting fascial tissue and minimize artefacts of fluid application. PMID:27609040

  12. Functional in vitro tension measurements of fascial tissue - a novel modified superfusion approach.

    Science.gov (United States)

    Schleip, R; Klingler, W; Wearing, S; Naylor, I; Zuegel, M; Hoppe, K

    2016-09-07

    While two laboratory techniques are commonly used to assess the tensile properties of muscle tissue, emerging evidence suggests that the fascial components of these tissues also serve an active role in force generation. Hence, we investigated whether these techniques are sensitive for assessment of fascial micromechanics. Force measurements on dissected fascial tissue were performed either using the classical immersion organ bath or using an improved superfusion approach simulating pulsed pharmacological triggers. Rat deep dorsal fascial strips as well as rat testicular capsule were pharmacologically challenged either with mepyramine or oxytocin. The classical immersion technique yielded a lower force response to mepyramine than the superfusion method (median: 367.4 vs. 555.4µN/mm(2)). Pause in irrigation before application reduced irregularities during bolus application. The superfusion approach was improved further by the following points: The high sensitivity of the superfusion method to bolus addition was voided by deviation of fluid supply during bolus addition. Although both methods demonstrated pharmacologically induced contractile responses in lumbar fascia samples, the modified superfusion method may improve force registrations of slow contracting fascial tissue and minimize artefacts of fluid application.

  13. New noninvasive approach assessing in vivo sun protection factor (SPF) using diffuse reflectance spectroscopy (DRS) and in vitro transmission.

    Science.gov (United States)

    Ruvolo Junior, Eduardo; Kollias, Nikiforos; Cole, Curtis

    2014-08-01

    absorbance matched to the UVA absorbance at 340 nm to complete the full spectral absorbance from which an estimate the SPF of the product can be calculated. Seventeen test materials with known in vivo SPF values were tested. Two of the tested products were PVC sunscreen thin films with 10-15 micrometers thickness and were used to investigate the absorption spectrum of these films when applied on different reflectance surfaces. Similar to the human in vivo SPF test, the developed methodology suggests limiting the use on Fitzpatrick skin phototypes I to III. The correlation of this new method with in vivo clinical SPF values was 0.98 (r2) with a slope of 1.007. This new methodology provides a new approach to determine SPF values without the extensive UV irradiation procedures (and biological responses) currently used to establish sunscreen efficacy. Further work will be conducted to establish methods for evaluation of products that are not photostable. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. In vitro investigation of cytochrome P450-mediated metabolism of dietary flavonoids

    DEFF Research Database (Denmark)

    Breinholt, Vibeke; Offord, E.A.; Brouwer, C.

    2002-01-01

    Human and mouse liver microsomes And membranes isolated from Escherichia coli, which expressed cytochrome P450 (CYP) 1A2, 3A4 2C9 or 2D6, were used to investigate CYP-mediated metabolism of five selected dietary flavonoids. In human and mouse liver microsomes kaempferol, apigenin and naringenin...... were hydroxylated at the 3'-position to yield their corresponding analogs quercetin, luteolin and eriodietyol, whereas hesperetin and tamarixetin were demethylated at the 4'-position to yield eriodictyol and quercetin. respectively, Microsomal flavonoid metabolism as potently inhibited by the CYP1A2...... inhibitors. fluvoxamine and alpha-naphthoflavone. Recombinant CYP1A2 as capable of metabolizing all five investigated flavonoids. CYP3A4 recombinant protein did not catalyze hesperetin demethylation. but showed similar metabolic profiles for the remaining compounds, as did human microsomes and recombinant...

  15. Phytochemical investigation GC-MS analysis and in vitro antimicrobial activity of Coleus forskohlii

    Directory of Open Access Journals (Sweden)

    Krishnamoorthy Rajkumar

    2015-12-01

    Full Text Available The aim of this study was to investigate the phytochemical constituents, gas chromatography-mass spectrometry (GC-MS analysis and antimicrobial activity of Coleus forskohlii. The different solvents such as ethanol, chloroform, acetone and aqueous extracts were identified pharmacologically as important bioactive compounds and their antimicrobial properties were studied. In the phytochemical investigation almost all the ethanol extract of leaf, stem and root having secondary metabolites like alkaloids, flavonoids, tannins, saponins, terpenoids, and steroids. The active constituents of the ethanol extract of C. forskohlii root was studied by GC-MS analysis. According to the antimicrobial results ethanol extract of C. froshkolii root showed highest antibacterial activity compared with stem and leaf. The highest antimicrobial activity was observed against Klebsiella pneumonia (19 mm and Candida albicans (16 mm in ethanol extract of root. Among the above extracts of leaf, stem and root, ethanol extract of root having antimicrobial activities due to the presence of phytoconstituents.

  16. A tiered approach for integrating exposure and dosimetry with in vitro dose-response data in the modern risk assessment paradigm

    Science.gov (United States)

    High-throughput (HT) risk screening approaches apply in vitro dose-response data to estimate potential health risks that arise from exposure to chemicals. However, much uncertainty is inherent in relating bioactivities observed in an in vitro system to the perturbations of biolog...

  17. Development of a combined in Vitro Physiologically Based Kinetic (PBK) and Monte Carlo modelling approach to predict interindividual human variation in phenol-induced developmental toxicity

    NARCIS (Netherlands)

    Strikwold, Marije; Spenkelink, Bert; Woutersen, Ruud A.; Rietjens, Ivonne M.C.M.; Punt, Ans

    2017-01-01

    With our recently developed in vitro physiologically based kinetic (PBK) modelling approach, we could extrapolate in vitro toxicity data to human toxicity values applying PBK-based reverse dosimetry. Ideally information on kinetic differences among human individuals within a population should be

  18. In Vitro and In Vivo Investigation of the Potential of Amorphous Microporous Silica as a Protein Delivery Vehicle

    Directory of Open Access Journals (Sweden)

    Amol Chaudhari

    2013-01-01

    Full Text Available Delivering growth factors (GFs at bone/implant interface needs to be optimized to achieve faster osseointegration. Amorphous microporous silica (AMS has a potential to be used as a carrier and delivery platform for GFs. In this work, adsorption (loading and release (delivery mechanism of a model protein, bovine serum albumin (BSA, from AMS was investigated in vitro as well as in vivo. In general, strong BSA adsorption to AMS was observed. The interaction was stronger at lower pH owing to favorable electrostatic interaction. In vitro evaluation of BSA release revealed a peculiar release profile, involving a burst release followed by a 6 h period without appreciable BSA release and a further slower release later. Experimental data supporting this observation are discussed. Apart from understanding protein/biomaterial (BSA/AMS interaction, determination of in vivo protein release is an essential aspect of the evaluation of a protein delivery system. In this regard micropositron emission tomography (μ-PET was used in an exploratory experiment to determine in vivo BSA release profile from AMS. Results suggest stronger in vivo retention of BSA when adsorbed on AMS. This study highlights the possible use of AMS as a controlled protein delivery platform which may facilitate osseointegration.

  19. New silver(I) complex with diazafluorene based ligand: Synthesis, characterization, investigation of in vitro DNA binding and antimicrobial studies

    Science.gov (United States)

    Movahedi, Elaheh; Rezvani, Ali Reza

    2017-07-01

    A novel diazafluorene based complex with silver, [Ag(dian)2 ] NO3 , where dian is N-(4,5-diazafluoren-9-ylidene)aniline, has been prepared and characterized by elemental analysis, IR spectroscopy, 1HNMR, UV-Vis spectroscopy and cyclic voltammetry. In order to explore the relationship between the structure and biological properties, DNA binding propensity and in vitro antibacterial property have also been studied. The mode of DNA-complex interaction has been investigated by electronic absorption titration, luminescence titration, competitive binding experiment, effect of ionic strength, thermodynamic studies, viscometric evaluation, circular dichroism spectroscopy and cyclic voltammetry. The results reveal that the complex binds to CT-DNA in a moderate intercalation capability with the partial insertion of a planar dian ligand between the base stacks of double-stranded DNA with binding constant (Kb) of 2.4 × 105 M-1. The viscosities and CD spectra of the DNA provide strong evidence for the intercalation. An in vitro antibacterial efficacy of the Ag(I) complex on a series of Gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) indicates that the complex exhibits a marked antibacterial activity. The minimum inhibitory concentrations of the complex indicate that it exhibits much higher antibacterial effect on standard bacterial strains of Escherichia coli and Staphylococcus aureus than those of silver nitrate, silver sulfadiazine. The bacterial inhibitions of the silver(I) complex are closely agreed to its DNA binding affinities.

  20. Rapid Prototyping for In Vitro Knee Rig Investigations of Prosthetized Knee Biomechanics: Comparison with Cobalt-Chromium Alloy Implant Material

    Directory of Open Access Journals (Sweden)

    Christian Schröder

    2015-01-01

    Full Text Available Retropatellar complications after total knee arthroplasty (TKA such as anterior knee pain and subluxations might be related to altered patellofemoral biomechanics, in particular to trochlear design and femorotibial joint positioning. A method was developed to test femorotibial and patellofemoral joint modifications separately with 3D-rapid prototyped components for in vitro tests, but material differences may further influence results. This pilot study aims at validating the use of prostheses made of photopolymerized rapid prototype material (RPM by measuring the sliding friction with a ring-on-disc setup as well as knee kinematics and retropatellar pressure on a knee rig. Cobalt-chromium alloy (standard prosthesis material, SPM prostheses served as validation standard. Friction coefficients between these materials and polytetrafluoroethylene (PTFE were additionally tested as this latter material is commonly used to protect pressure sensors in experiments. No statistical differences were found between friction coefficients of both materials to PTFE. UHMWPE shows higher friction coefficient at low axial loads for RPM, a difference that disappears at higher load. No measurable statistical differences were found in knee kinematics and retropatellar pressure distribution. This suggests that using polymer prototypes may be a valid alternative to original components for in vitro TKA studies and future investigations on knee biomechanics.

  1. New approach for a reliable in vitro sun protection factor method Part I: Principle and mathematical aspects.

    Science.gov (United States)

    Miksa, S; Lutz, D; Guy, C

    2015-12-01

    Development of an international harmonized in vitro method for sun protection factor (SPF) assessment is currently in progress because of the lack of both reproducibility and accuracy in the current methodology. The aim of this article was to focus on the principle and mathematical aspects (Part I) of a new approach for a reproducible and correlated in vitro test method to obtain results similar to the in vivo SPF for labelling purposes. The currently used in vitro test is based on the spectroradiometric measurement of the residual ultraviolet (UV) transmitted through a thin layer of sunscreen spread on a substrate. To reach the goals of reproducibility and accuracy, the specifications of the key parameters and different steps of the procedure are clearly described in this study. Once reproducibility is obtained with an ad hoc procedure, the accuracy of the SPF values for a large number of products can be demonstrated with the prerequisite of a single UV irradiation dose and a multisubstrate solution. Using a total of 27 samples, the mean coefficient of variation was found to be vitro SPF assessment that can be closely correlated to the in vivo SPF for labelling purposes. The second part study will focus on the practical aspects and implementation (Part II) achieved using the present method, will validate the robustness of the models and demonstrate the need to have different product categories to reach a reliable in vitro SPF method adaptable for all products available in the market. © 2015 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  2. A Proteomic Approach for Investigation of Bee Products: Royal Jelly, Propolis and Honey

    Directory of Open Access Journals (Sweden)

    Peter Raspor

    2012-01-01

    Full Text Available Bee products such as royal jelly, honey and propolis have been reported to possess several biological activities. In order to better understand their mechanism of action and, consequently, their efficiency and safety, 'omic' approaches are used. Here cases with proteomic approach are indicated. In addition to studying biological activity at a proteome level, a proteomic approach for investigation of bee products has also been applied in analyzing proteins as their (bioactive components.

  3. Exploring Game Transfer Phenomena: a multimodal research approach for investigating video games' effects

    OpenAIRE

    Ortiz de Gortari, AB

    2015-01-01

    Video games are evolving and are becoming ever more immersive. Consequently, it is necessary to understand their effects on gamers’ psychological wellbeing. The impact on cognition, affect and behaviour has mostly been investigated separately and sometimes from narrow approaches that limit the understanding of the video games’ effects. This thesis investigates the effects of playing video games from a novel, multimodal and broad research approach that is termed "Game Transfer Phenomena" (GTP)...

  4. Investigation of the bystander effect in MRC5 cells after acute and fractionated irradiation in vitro

    Directory of Open Access Journals (Sweden)

    Shokouhozaman Soleymanifard

    2014-01-01

    Full Text Available Radiation-induced bystander effect (RIBE has been defined as radiation responses observed in nonirradiated cells. It has been the focus of investigators worldwide due to the deleterious effects it induces in nonirradiated cells. The present study was performed to investigate whether acute or fractionated irradiation will evoke a differential bystander response in MRC5 cells. A normal human cell line (MRC5, and a human lung tumor cell line (QU-DB were exposed to 0, 1, 2, and 4Gy of single acute or fractionated irradiation of equal fractions with a gap of 6 h. The MRC5 cells were supplemented with the media of irradiated cells and their micronucleus frequency was determined. The micronucleus frequency after single and fractionated irradiation did not vary significantly in the MRC5 cells conditioned with autologous or QU-DB cell-irradiated media, except for 4Gy where the frequency of micronucleated cells was lower in those MRC5 cells cultured in the media of QU-DB-exposed with a single dose of 4Gy. Our study demonstrates that the radiation-induced bystander effect was almost similar after single acute and fractionated exposure in MRC5 cells.

  5. Carboxymethyl guar gum nanoparticles for drug delivery applications: Preparation and preliminary in-vitro investigations

    Energy Technology Data Exchange (ETDEWEB)

    Dodi, G., E-mail: gianina.dodi@yahoo.co.uk [“Gheorghe Asachi” Technical University of Iasi (Romania); SCIENT — Research Centre for Instrumental Analysis, Bucharest (Romania); Pala, A. [University of Sassari, Sassari (Italy); Barbu, E. [University of Portsmouth, Portsmouth (United Kingdom); Peptanariu, D. [“Petru Poni” Institute of Macromolecular Chemistry, Iasi (Romania); Hritcu, D.; Popa, M.I. [“Gheorghe Asachi” Technical University of Iasi (Romania); Tamba, B.I. [“Gr. T. Popa” University of Medicine and Pharmacy, Iasi (Romania)

    2016-06-01

    Carboxymethyl guar gum (CMGG) synthesized from commercially available polysaccharide was formulated into nanoparticles via ionic gelation using trisodium trimetaphosphate (STMP) as cross-linking agent. Characterisation using a range of analytical techniques (FTIR, NMR, GPC, TGA and DLS) confirmed the CMGG structure and revealed the effect of the CMGG and STMP concentration on the main characteristics of the obtained nanoformulations. The average nanoparticle diameter was found to be around 208 nm, as determined by dynamic light scattering (DLS) and confirmed by scanning electron microscopy (SEM) and nanoparticle tracking analysis (NTA). Experiments using simulated gastric and intestinal fluids evidenced significant pH-dependent drug release behaviour of the nanoformulations loaded with Rhodamine B (RhB) as a model drug (loading capacity in excess of 83%), as monitored by UV–Vis. While dose-dependent cytotoxicity was observed, the nanoformulations appeared completely non-toxic at concentrations below 0.3 mg/mL. Results obtained so far suggest that carboxymethylated guar gum nanoparticles formulated with STMP warrant further investigations as polysaccharide based biocompatible drug nanocarriers. - Highlights: • Carboxymethyl guar gum nanoparticles preparation by ionic gelation • The optimum synthesis system designed particles around 200 nm • The nanoformulations appeared completely non-toxic at specific concentrations • The loaded formulations evidenced significant pH-dependent drug release behaviour • The results encourage further investigations as polysaccharidic drug nanocarriers.

  6. Investigation of the bystander effect in MRC5 cells after acute and fractionated irradiation in vitro.

    Science.gov (United States)

    Soleymanifard, Shokouhozaman; Toossi, Mohammad Taghi Bahreyni; Samani, Roghayeh Kamran; Mohebbi, Shokoufeh

    2014-04-01

    Radiation-induced bystander effect (RIBE) has been defined as radiation responses observed in nonirradiated cells. It has been the focus of investigators worldwide due to the deleterious effects it induces in nonirradiated cells. The present study was performed to investigate whether acute or fractionated irradiation will evoke a differential bystander response in MRC5 cells. A normal human cell line (MRC5), and a human lung tumor cell line (QU-DB) were exposed to 0, 1, 2, and 4Gy of single acute or fractionated irradiation of equal fractions with a gap of 6 h. The MRC5 cells were supplemented with the media of irradiated cells and their micronucleus frequency was determined. The micronucleus frequency after single and fractionated irradiation did not vary significantly in the MRC5 cells conditioned with autologous or QU-DB cell-irradiated media, except for 4Gy where the frequency of micronucleated cells was lower in those MRC5 cells cultured in the media of QU-DB-exposed with a single dose of 4Gy. Our study demonstrates that the radiation-induced bystander effect was almost similar after single acute and fractionated exposure in MRC5 cells.

  7. Investigating Trait Emotional Intelligence among School Leaders: Demonstrating a Useful Self-Assessment Approach

    Science.gov (United States)

    Benson, Robert; Fearon, Colm; McLaughlin, Heather; Garratt, Sara

    2014-01-01

    An exploratory study of two grammar schools in the South East of England is used to justify and demonstrate a self-assessed approach that investigates "trait" emotional intelligence (EI) among school leaders. First, the theoretical underpinnings of "ability" and "trait" EI approaches are critically compared based on…

  8. Investigating the joint development of approach bias and adolescent alcohol use

    NARCIS (Netherlands)

    Janssen, T.; Wood, M.; Larsen, H.; Peeters, M.; Vollebergh, W.A.M.; Wiers, Reinout W

    2015-01-01

    Background We investigated the joint development between implicit approach bias and early adolescent alcohol use, and examined whether the link between approach bias and alcohol use was moderated by working memory (WM). Methods The current study used data from a 2-year, 4-wave online sample of 378

  9. Investigating How Digital Technologies Can Support a Triad-Approach for Student Assessment in Higher Education

    Science.gov (United States)

    Vaughan, Norman

    2013-01-01

    The purpose of this research study was to investigate if and how digital technologies could be used to support a triad-approach for student assessment in higher education. This triad-approach consisted of self-reflection, peer feedback, and instructor assessment practices in a pre-service teacher education course at a Canadian university. Through…

  10. Prediction of pharmacokinetic and toxicological parameters of a 4-phenylcoumarin isolated from geopropolis: In silico and in vitro approaches.

    Science.gov (United States)

    da Cunha, Marcos Guilherme; Franco, Gilson César Nobre; Franchin, Marcelo; Beutler, John A; de Alencar, Severino Matias; Ikegaki, Masaharu; Rosalen, Pedro Luiz

    2016-11-30

    In silico and in vitro methodologies have been used as important tools in the drug discovery process, including from natural sources. The aim of this study was to predict pharmacokinetic and toxicity (ADME/Tox) properties of a coumarin isolated from geopropolis using in silico and in vitro approaches. Cinnamoyloxy-mammeisin (CNM) isolated from Brazilian M. scutellaris geopropolis was evaluated for its pharmacokinetic parameters by in silico models (ACD/Percepta™ and MetaDrug™ software). Genotoxicity was assessed by in vitro DNA damage signaling PCR array. CNM did not pass all parameters of Lipinski's rule of five, with a predicted low oral bioavailability and high plasma protein binding, but with good predicted blood brain barrier penetration. CNM was predicted to show low affinity to cytochrome P450 family members. Furthermore, the predicted Ames test indicated potential mutagenicity of CNM. Also, the probability of toxicity for organs and tissues was classified as moderate and high for liver and kidney, and moderate and low for skin and eye irritation, respectively. The PCR array analysis showed that CNM significantly upregulated about 7% of all DNA damage-related genes. By exploring the biological function of these genes, it was found that the predicted CNM genotoxicity is likely to be mediated by apoptosis. The predicted ADME/Tox profile suggests that external use of CNM may be preferable to systemic exposure, while its genotoxicity was characterized by the upregulation of apoptosis-related genes after treatment. The combined use of in silico and in vitro approaches to evaluate these parameters generated useful hypotheses to guide further preclinical studies. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  11. In vitro investigations of Cynara scolymus L. extract on cell physiology of HepG2 liver cells

    Directory of Open Access Journals (Sweden)

    Gesine Löhr

    2009-06-01

    Full Text Available The objective of this study was the investigation of a potential influence of artichoke leaf extract (ALE on the cell physiology and gene expression of phase I/II enzymes of human liver cells HepG2 and investigation on potential cell protective effects against ethanol-induced cell toxicity against HepG2 cells. Cell biological assays under in vitro conditions using HepG2 liver cells and investigation of mitochondrial activity (MTT test, proliferation assay (BrdU incorporation ELISA, LDH as toxicity marker, gene expression analysis by RT-PCR and enzyme activity of glutationtransferase. Artichocke extract, containing 27% caffeoylquinic acids and 7% flavonoids induced mitochondrial activity, proliferation and total protein content under in vitro conditions in human liver cells HepG2. These effects could not be correlated to the well-known artichoke secondary compounds cynarin, caffeic acid, chlorogenic acid, luteolin and luteolin-7-O-glucoside. The flavones luteolin and luteolin-7-O-glucoside had inhibitory effects at 100 µg/mL level on HepG2 cells, with luteolin being a significant stronger inhibitor compared to the respective glucoside. Artichoke leaf extract had minor stimulating effect on gene expression of CYP1A2, while CYP3A4, GGT, GPX2, GSR and GST were slightly inhibited. GST inhibition under in vitro conditions was also shown by quantification of GST enzyme activity. Induction of gene expression of CYP1A2 was shown to be supraadditive after simultaneous application of ethanol plus artichoke extract. Artichoke leaf extract exhibited cell protective effects against ethanol-induced toxicity within cotreatment under in vitro conditions. Also H2O2 damage was significantly inhibited by simultaneous artichoke incubation. Pre- and posttreatments did not exert protective effects. DMSO-induced toxicity was significantly reduced by pre-, post- and cotreatment with artichoke extract and especially with luteolin-7-O-glucoside, indicating a direct

  12. In vitro investigations of repulsion during laser lithotripsy using a pendulum set-up.

    Science.gov (United States)

    Sroka, Ronald; Haseke, Nicolas; Pongratz, Thomas; Hecht, Volkmar; Tilki, Derya; Stief, Christian G; Bader, Markus Jürgen

    2012-05-01

    Ureteroscopic laser lithotripsy is a commonly used technique to treat ureteral calculi.The type of energy source used is one of the main influences of retrograd calculi propulsion. Using a momentum pendulum under-water set-up the induced momentum and the initial velocity were investigated. Pulsed laser light from three different clinically available laser systems, including a Ho:YAG laser, a frequency-doubled double-pulse (second harmonic generation, SHG) Nd:YAG laser and a flash-lamp pumped dye (FLPD) laser, were transmitted via flexible fibres of different core diameter to the front of the pendulum sinker. Single pulses at variable pulse energy, according to the clinical laser parameter settings, were applied to the target sinker, thus causing a repulsion-induced deflection which was documented by video recording. The maximum deflection was determined. Solving the differential equation of a pendulum gives the initial velocity, the laser-induced momentum and the efficiency of momentum transfer. The induced deflection as well as the starting velocity of the two short-duration pulsed laser systems (SHG Nd:YAG, FLPD) were similar (s (max) = 2-3.6 cm and v (0) = 150-200 mm/s, respectively), whereas both values were lower using the Ho:YAG laser with a long pulse duration (s (max) = 0.9--1.6 cm and v (0) = 60-105 mm/s, respectively). The momentum I induced by the Ho:YAG laser was only 50% and its transfer efficacy η (Repuls) was reduced to less than 5% of the values of the two short-pulsed laser systems. This investigation clearly showed the variable parts and amounts of repulsion using different pulsed lasers in an objective and reproducible manner. The momentum transfer efficiency could be determined without any physical friction problems. Further investigations are needed to compare stone fragmentation techniques with respect to laser repulsion and its clinical impact.

  13. Investigation of in vitro digestibility of dietary microalga Chlorella vulgaris and cyanobacterium Spirulina platensis as a nutritional supplement.

    Science.gov (United States)

    Kose, Ayse; Ozen, Mehmet O; Elibol, Murat; Oncel, Suphi S

    2017-07-01

    Microalgal proteins are promising sources for functional nutrition and a sustainable candidate for nutraceutical formulations. They also gain importance due to emerging focus on a healthy nutrition and increase in the number of chronic diseases. In this study, dried dietary species of microalga, Chlorella vulgaris, and cyanobacterium Spirulina platensis were hydrolyzed with pancreatin enzyme to obtain protein hydrolysates. The hydrolysis yield of biomass was 55.1 ± 0.1 and 64.8 ± 3.6% for C. vulgaris and S. platensis; respectively. Digestibility, as an indicator for dietary utilization, was also investigated. In vitro protein digestibility (IVPD) values depicted that cell wall structure due to the taxonomical differences affected both hydrolysis and digestibility yield of the crude biomass (p microalgae, which shows elevated digestibility values as a sustainable and reliable source.

  14. In vitro investigation of efficient photodynamic therapy using a nonviral vector; hemagglutinating virus of Japan envelope

    Science.gov (United States)

    Sakai, Makoto; Fujimoto, Naohiro; Ishii, Katsunori; Nakamura, Hiroyuki; Kaneda, Yasufumi; Awazu, Kunio

    2012-07-01

    Photodynamic therapy (PDT) is a photochemical modality approved for cancer treatment. PDT has demonstrated efficacy in early stage lung cancer and esophageal cancer. The accumulation of photosensitizers in cancer cells is necessary to enhance the therapeutic benefits of PDT; however, photosensitizers have low uptake efficiency. To overcome this limitation, a drug delivery system, such as the hemagglutinating virus of Japan envelope (HVJ-E) vector, is required. In this study, the combination of PDT and HVJ-E was investigated for enhancing the efficacy of PDT. The photosensitizers that were evaluated included 5-aminolaevulinic acid (5-ALA), protoporphyrin IX (PPIX), and HVJ-PPIX. The uptake of the photosensitizers as increased twenty-fold with the addition of HVJ-E. The cytotoxicity of conventional 5-ALA was enhanced by the addition of HVJ-E vector. In conclusion, HVJ-E vector improved the uptake of photosensitizers and the PDT effect.

  15. Nanostructured bioglass thin films synthesized by pulsed laser deposition: CSLM, FTIR investigations and in vitro biotests

    Science.gov (United States)

    Floroian, L.; Savu, B.; Stanciu, G.; Popescu, A. C.; Sima, F.; Mihailescu, I. N.; Mustata, R.; Sima, L. E.; Petrescu, S. M.; Tanaskovic, D.; Janackovic, Dj.

    2008-12-01

    We report the synthesis by pulsed laser deposition of thin structures of two bioactive glasses belonging to the SiO 2-Na 2O-K 2O-CaO-MgO-P 2O 5 system, on medical grade Ti substrates. We evaluated their biocompatibility after immersion in simulated body fluids and by performing cells adhesion tests. The films were characterized by confocal scanning laser microscopy and Fourier transform infrared spectrometry, before and after 30 and 46 days immersion in fluids. Our studies demonstrated that deposited coatings were degraded in simulated fluids. A new apatite layer was synthesized by ions changing with the fluid during the decomposition of bioglasses. We investigated after immersion in fluids cells adhesion and the cytoskelet organization of synthesized structures, by fluorescence microscopy. A good adhesion to bioglass coatings was evidenced.

  16. Investigations on in vitro anti-carcinogenic potential of L-carnosine in liver cancer cells.

    Science.gov (United States)

    Ding, Minghui; Jiao, Guihua; Shi, Haizhou; Chen, Yanrong

    2017-07-27

    This study was carried out to investigate the anti-carcinogenic effect of L-carnosine in human carcinoma cells (SNU-423). The SNU-423 cancer cells were cultured at a density of 2 × 104 cells/well in Dulbecco modified Eagle medium. After 24 h of adherence, the cells were treated with L-carnosine (0.2 and 1 mg/mL) for 48 h. Then, cell viability was assessed by sulforhodamine assay, while mitochondrial dysfunction was measured by fluorescence microscopy using chromatin-specific dye Hoechst 33258. Intracellular levels of ROS were assayed by fluorescence spectroscopy with 2',7'-dichlorofluorescein diacetate (DCFDA). L-Carnosine significantly inhibited the growth of the SNU-423 cells (p carcinogenic effects in human liver cancer cells.

  17. Investigation of biomaterials by human epithelial gingiva cells: an in vitro study

    Directory of Open Access Journals (Sweden)

    Neunzehn Jörg

    2012-12-01

    Full Text Available Abstract Introduction In modern medicine and dentistry the use of biomaterials is a fast developing field of increasing interest. Especially in dentistry the interaction between biomaterials like implant materials and the soft tissue in the oral cavity is in the focus of daily research. In this context the high importance of testing materials and their surfaces concerning their biocompatibility towards corresponding cells is very likely. For this purpose this study investigates cells derived from human gingival biopsies on different materials and surfaces. Methods Cells in this study were cultivated out of human biopsies by a grow out explant technique and were sub cultivated on titanium, zirconium dioxide and collagen membrane specimens. To characterise the cells on the material surfaces used in this study immunohistochemical and histological staining techniques as well as different methods of microscopy (light microscopy and SEM were applied. Results With the aid of the explant technique and the chosen cell cultivation method it was possible to investigate the human gingiva derived cells on different materials. The data of the present study show that the human gingival cells attach and proliferate on all three tested materials by exhibiting characteristic gingival keratinocyte protein expression even after long periods of culture e.g. up to 70 days. Conclusions It could be shown that the three tested materials titanium, zirconium dioxide and collagen membrane (and their special surfaces are good candidates for the application as materials in the dental gingival environment or, in the case of the collagen membrane as scaffold/cell-carrier for human gingival cells in tissue engineering.

  18. Binding of 8-methoxypsoralen to DNA in vitro: Monitoring by spectroscopic and chemometrics approaches

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Xiaoyue; Zhang, Guowen, E-mail: gwzhang@ncu.edu.cn; Wang, Langhong

    2014-10-15

    8-Methoxypsoralen (8-MOP) is a naturally occurring furanocoumarin with a variety of biological and pharmacological activities. The binding mechanism of 8-MOP to calf thymus DNA (ctDNA) at physiological pH was investigated by multi-spectroscopic techniques including UV–vis absorption, fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy along with DNA melting studies and viscosity measurements. The multivariate curve resolution-alternating least squares (MCR-ALS) chemometrics approach was introduced to resolve the expanded UV–vis spectral data matrix, and both the pure spectra and the equilibrium concentration profiles for the components (8-MOP, ctDNA and 8-MOP-ctDNA complex) in the system were successfully obtained to monitor the 8-MOP-ctDNA interaction. The results suggested that 8-MOP could bind to ctDNA via intercalation binding as evidenced by significant increases in melting and relative viscosity of ctDNA and competitive study using acridine orange (AO) as a fluorescence probe. The positive values of enthalpy and entropy change suggested that hydrogen bonds and van der Waals forces played a predominant role in the binding process. Further, FT-IR and CD spectra analysis indicated that 8-MOP preferentially bound to A–T base pairs with no major perturbation in ctDNA double helix conformation. Moreover, molecular docking was employed to exhibit the specific binding mode of 8-MOP to ctDNA intuitively. - Highlights: • The interaction processes of 8-MOP with ctDNA was monitored by MCR-ALS approach. • The binding mode of 8-MOP to ctDNA was an intercalation. • 8-MOP most likely bound to adenine and thymine base pairs of ctDNA. • Molecular docking illustrated the specific binding.

  19. Investigating the biological impacts of nanoengineered materials in Caenorhabditis elegans and in vitro

    Science.gov (United States)

    Contreras, Elizabeth Quevedo

    In nematode Caenorhabditis elegans, the chronic and multi-generational toxicological effects of commercially relevant engineered nanoparticles (ENPs), such as quantum dots (QDs) and silver (AgNP) caused significant changes in a number of physiological endpoints. The increased water-solubility of ENPs in commercial products, for example, makes them increasingly bioavailable to terrestrial organisms exposed to pollution and waste in the soil. Since 2008, attention to the toxicology of nanomaterials in C. elegans continues to grow. Quantitative data on multiple physiological endpoints paired with metal analysis show the uptake of QDs and AgNPs, and their effects on nematode fitness. First, C. elegans were exposed for four generations through feeding to amphiphilic polymer coated CdSe/ZnS (core-shell QDs), CdSe (core QDs), and different sizes of AgNPs. These ENPs were readily ingested. QDs were qualitatively imaged in the digestive tract using a fluorescence microscopy and their and AgNP uptake quantitatively measured using ICP-MS. Each generation was analyzed for changes in lifespan, reproduction, growth and motility using an automated computer vision system. Core-shell QDs had little impact on C. elegans due to its metal shell coating. In contrast, core QDs lacked a metal shell coating, which caused significant changes to nematode physiology. iii In the same way, at high concentrations of 100 ppm, AgNP caused the most adverse effect to lifespan and reproduction related to particle size, but its adverse effect to motility had no correlation to particle size. Using C. elegans as an animal model allowed for a better understanding of the negative impacts of ENPs than with cytotoxicity tests. Lastly, to test the toxicity of water-dispersed fullerene (nanoC60) using human dermal fibroblast cells, this thesis investigated a suite of assays and methods in order to establish a standard set of cytotoxicity tests. Ten assays and methods assessed nanoC60 samples of different

  20. An in vitro investigation of indigenous South African medicinal plants used to treat oral infections.

    Science.gov (United States)

    Akhalwaya, S; van Vuuren, S; Patel, M

    2018-01-10

    Over a 120 South African medicinal plants are used for the treatment of oral diseases. Despite the vast collection of antimicrobial studies being done on South African plants, there is still limited research on pathogens associated with oral infections. In consultation with the available ethnobotanical literature, this study investigates the antimicrobial efficacy of some South African medicinal plants against oral pathogens. To provide a detailed account of the antimicrobial properties of selected South African medicinal plants used traditionally to treat oral infections. The effect on Streptococcus mutans biofilm formation and the toxicity profiles of these plants are also investigated. A total of 136 aqueous and organic extracts and six essential oils were prepared from 31 different plant species. These plant samples were screened for antimicrobial efficacy against nine oral pathogens using the micro-titre plate dilution assay. Plant extracts that were found to have noteworthy antimicrobial activity against S. mutans were further evaluated on the effect on S. mutans biofilm formation using the glass slide technique. The toxicity profiles of plant samples that were found to have noteworthy antimicrobial activity were evaluated using the brine shrimp lethality assay. The organic extract of Cissampelos torulosa stems displayed the lowest MIC value of 0.05mg/mL against both Lactobacillus spp. This high antimicrobial activity was also observed with the organic extract of Spirostachys africana leaves against Candida albicans. In some instances, a direct relationship was found between the traditional use of the plant and the antimicrobial activity observed. For example, noteworthy activity (MIC plant traditionally used to treat oral thrush. Englerophytum magalismonatanum stems displayed notable activity against both Streptococcus spp. (MIC 0.83mg/mL against S. mutans and MIC 0.67mg/mL against S. sanguis). Spirostachys africana leaves displayed the greatest anti

  1. Investigation of Microbubble Cavitation-Induced Calcein Release from Cells In Vitro.

    Science.gov (United States)

    Maciulevičius, Martynas; Tamošiūnas, Mindaugas; Jakštys, Baltramiejus; Jurkonis, Rytis; Venslauskas, Mindaugas Saulius; Šatkauskas, Saulius

    2016-12-01

    In the present study, microbubble (MB) cavitation signal analysis was performed together with calcein release evaluation in both pressure and exposure duration domains of the acoustic field. A passive cavitation detection system was used to simultaneously measure MB scattering and attenuation signals for subsequent extraction efficiency relative to MB cavitation activity. The results indicate that the decrease in the efficiency of extraction of calcein molecules from Chinese hamster ovary cells, as well as cell viability, is associated with MB cavitation activity and can be accurately predicted using inertial cavitation doses up to 0.18 V × s (R 2  > 0.9, p cavitation in the sono-extraction process. To our knowledge, this study is the first to (i) investigate small molecule extraction from cells via sonoporation and (ii) relate the extraction process to the quantitative characteristics of MB cavitation acoustic spectra. Copyright © 2016 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  2. An in vitro investigation of wear resistance and hardness of composite resins.

    Science.gov (United States)

    Cao, Liqun; Zhao, Xinyi; Gong, Xu; Zhao, Shouliang

    2013-01-01

    The aim of the present study was to investigate the wear resistance and hardness of five kinds of composite resins. Sixty-five specimens were fabricated with one nano-hybrid (Charisma Diamond), two micro-hybrid (3MZ250, Clearfil AP-X) and two packable (3MP60, Surefil) composite resins, according to a randomized complete block design (n=13, 8 for wear test; 5 for hardness test). The composites were filled in a rectangular mold, and light polymerization. After storage in 37°C deionized water for 24h, all specimens were tested with a custom-made toothbrush machine with a stainless-steel ball as antagonist (3N loads, 1Hz, 6×10(5) cycles) immersed in calcium fluoride slurry. Wear volume, hardness and surface structure of each tested material was examined by a three-dimensional non-contact optical profilometer, Vickers indentation technique and scanning electron microscope. The volume loss ranked from least to most as follows: Charisma Diamond, P60, Z250, Clearfil AP-X and Surefil. Regarding hardness, the rank from highest to lowest as follows: Clearfil AP-X, P60, Surefil, Z250, Charisma Diamond. The interactions between wear resistance and microhardness were not significant. The custom-made machine is considered suitable to simulate sliding of an antagonist cusp on an opposing occlusal composite restoration. Nanofilled composite may have superior wear compared to other composite resins.

  3. Investigations of plant-derived products with the in vitro comet assay

    Directory of Open Access Journals (Sweden)

    Luc Verschaeve

    2015-08-01

    It is impossible to perform a wide range of tests for screening purposes and often only the Ames assay is performed, which is insufficient. Furthermore, this test is most probably not the best choice when plant extracts need to be tested, because they often contain high amounts of histidine and have antibacterial properties. We have participated in many screening programs of medicinal plants and used different genotoxicity tests (mainly Ames assay, Vitotox test, micronucleus test and comet assay. Or results revealed that a combination of the Vitotox test and comet assay provides sufficiently reliable data with respect to genotoxicity as well as antigenotoxicity. This holds true for the testing of (medicinal plant extracts but also other plant derived products, for example those aimed at identifying novel TB chemotherapeutic drugs. The investigation of smoke and smoke compounds as enhancers of seed germination and smoke treated plants provides another example in which the comet assay proved to be valuable in the assessment of potential adverse health effects resulting from such treatment.

  4. Chitosan nanoparticles for antimicrobial photodynamic inactivation: characterization and in vitro investigation.

    Science.gov (United States)

    Chen, Chueh-Pin; Chen, Chin-Tin; Tsai, Tsuimin

    2012-01-01

    The growing resistance to antibiotics has rendered antimicrobial photodynamic inactivation (PDI) an attractive alternative treatment modality for infectious diseases. Chitosan (CS) was shown to further potentiate the PDI effect of photosensitizers and was therefore used in this study to investigate its ability to potentiate the activity of erythrosine (ER) against bacteria and yeast. CS nanoparticles loaded with ER were prepared by ionic gelation method and tested for their PDI efficacy on planktonic cells and biofilms of Streptococcus mutans, Pseudomonas aeruginosa and Candida albicans. The nanoparticles were characterized for their size, polydispersity index and zeta potential. No toxicity was observed when planktonic cells and biofilms were treated with the nanoparticles in the dark. However, when the cells were exposed to light irradiation after treatment with free ER or ER/CS nanoparticles, a significant phototoxicity was observed. The antimicrobial activity of ER/CS nanoparticles was significantly higher than ER in free form. The particle size and incubation time of the nanoparticles also appeared to be important factors affecting their PDI activity against S. mutans and C. albicans. © 2012 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2012 The American Society of Photobiology.

  5. Investigation of hyfrecators and their in vitro interference with implantable cardiac devices.

    Science.gov (United States)

    Weyer, Christopher; Siegle, Ronald J; Eng, Guillaume Girard P

    2012-11-01

    Guidelines exist for minimizing potential electromagnetic interference (EMI) with electrosurgical equipment in patients with cardiac rhythm management (CRM) devices. These guidelines encompass all electrosurgical devices but are not specific for hyfrecators. To investigate the potential interference of CRM devices by hyfrecators. Using a collagen-based saline gel, three implantable pulse generators (pacemakers) and three implantable cardioverter defibrillators were tested to measure the EMI from two commonly used hyfrecators. The six devices were tested using the hyfrecator under normal use settings and on maximum power. Hyfrecators did not interfere with defibrillators and affected pacemakers only when used in close proximity to the device. For the pacemakers, atrial inhibition was observed at a distance of 3 cm on maximum hyfrecator settings and 1 cm at normal use settings. Ventricular inhibition occurred in very close proximity to the device (<1 cm) or in direct contact. Hyfrecators are safe to use in patients with defibrillators and can be used in pacemaker patients within 2 inches of the device perimeter. © 2012 by the American Society for Dermatologic Surgery, Inc. Published by Wiley Periodicals, Inc.

  6. An extensive cocktail approach for rapid risk assessment of in vitro CYP450 direct reversible inhibition by xenobiotic exposure.

    Science.gov (United States)

    Spaggiari, Dany; Daali, Youssef; Rudaz, Serge

    2016-07-01

    Acute exposure to environmental factors strongly affects the metabolic activity of cytochrome P450 (P450). As a consequence, the risk of interaction could be increased, modifying the clinical outcomes of a medication. Because toxic agents cannot be administered to humans for ethical reasons, in vitro approaches are therefore essential to evaluate their impact on P450 activities. In this work, an extensive cocktail mixture was developed and validated for in vitro P450 inhibition studies using human liver microsomes (HLM). The cocktail comprised eleven P450-specific probe substrates to simultaneously assess the activities of the following isoforms: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 2J2 and subfamily 3A. The high selectivity and sensitivity of the developed UHPLC-MS/MS method were critical for the success of this methodology, whose main advantages are: (i) the use of eleven probe substrates with minimized interactions, (ii) a low HLM concentration, (iii) fast incubation (5min) and (iv) the use of metabolic ratios as microsomal P450 activities markers. This cocktail approach was successfully validated by comparing the obtained IC50 values for model inhibitors with those generated with the conventional single probe methods. Accordingly, reliable inhibition values could be generated 10-fold faster using a 10-fold smaller amount of HLM compared to individual assays. This approach was applied to assess the P450 inhibition potential of widespread insecticides, namely, chlorpyrifos, fenitrothion, methylparathion and profenofos. In all cases, P450 2B6 was the most affected with IC50 values in the nanomolar range. For the first time, mixtures of these four insecticides incubated at low concentrations showed a cumulative inhibitory in vitro effect on P450 2B6. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Characterization of Pharmacokinetics in the Göttingen Minipig with Reference Human Drugs: An In Vitro and In Vivo Approach.

    Science.gov (United States)

    Lignet, Floriane; Sherbetjian, Eva; Kratochwil, Nicole; Jones, Russell; Suenderhauf, Claudia; Otteneder, Michael B; Singer, Thomas; Parrott, Neil

    2016-10-01

    This study aims to expand our understanding of the mechanisms of drug absorption, distribution, metabolism and excretion in the Göttingen minipig to aid a knowledge-driven selection of the optimal species for preclinical pharmaceutical research. The pharmacokinetics of seven reference compounds (antipyrine, atenolol, cimetidine, diazepam, hydrochlorothiazide, midazolam and theophylline) was investigated after intravenous and oral dosing in minipigs. Supportive in vitro data were generated on hepatocellularity, metabolic clearance in hepatocytes, blood cell and plasma protein binding and metabolism routes. Systemic plasma clearance for the seven drugs ranged from low (1.1 ml/min/kg, theophylline) to close to liver blood flow (37.4 ml/min/kg, cimetidine). Volume of distribution in minipigs ranged from 0.7 L/kg for antipyrine to 3.2 L/kg for hydrochlorothiazide. A gender-related difference of in vivo metabolic clearance was observed for antipyrine. The hepatocellularity for minipig was determined as 124 Mcells/g liver, similar to the values reported for human. Based on these data a preliminary in vitro to in vivo correlation (IVIVC) for metabolic clearance measured in hepatocytes was investigated. Metabolite profiles of diazepam and midazolam compared well between minipig and human. The results of the present study support the use of in vitro metabolism data for the evaluation of minipig in preclinical research and safety testing.

  8. In vitro investigation of the follicular penetration of porcine ear skin using a nanoparticle-emulsion containing the antiseptic polihexanide In vitro investigation of the follicular penetration of porcine ear skin

    Science.gov (United States)

    Ulmer, M.; Patzelt, A.; Vergou, T.; Lademann, J.; Richter, H.; Kramer, A.; Müller, G.; Sterry, W.; Lange-Asschenfeldt, B.

    2012-05-01

    Earlier investigations regarding the distribution of the bacterial flora on the human skin demonstrate that the hair follicle acts as a bacterial reservoir, providing a quick source for secondary recontamination. These findings highlight the importance of the hair follicle as a target for modern antiseptics. In the present study, we have assessed the follicular penetration of a curcumin-labeled particle-associated antiseptic into porcine skin by laser scanning microscopy. Therefore, the follicular penetration depth of the curcumin-labeled particle-associated antiseptic was compared to the follicular penetration depth of curcumin-labeled particles without antiseptic. The investigation was performed in vitro using porcine skin biopsies. By superposition of the images acquired in the transmission and the fluorescent modus, it was possible to visualize the distribution of the fluorescent dye inside the hair follicles. Quantitative and qualitative results showed that both dispersions penetrated efficiently into the hair follicles. The average penetration depth of the particles with attached antiseptic polihexanide was significantly higher than that of particles without the attached antiseptic. Also, whilst very little sample preparation was needed, laser scanning microscopy was found to be an efficient tool to visualize the skin relief and in particular the hair follicle shaft and localize fluorescent markers within the skin tissue and hair follicles.

  9. Recovery of the Zika virus through an in vitro ligation approach.

    Science.gov (United States)

    Deng, Cheng-Lin; Zhang, Qiu-Yan; Chen, Dong-Dong; Liu, Si-Qing; Qin, Cheng-Feng; Zhang, Bo; Ye, Han-Qing

    2017-07-01

    In this study, an in vitro ligation method was developed to assemble a full-length infectious cDNA clone of the Zika virus (ZIKV). Four contiguous cDNA subclones covering the complete ZIKV genome were constructed with unique BglI restriction sites at the ends of each fragment. The BglI restriction sites only allow in vitro ligation to happen between interconnecting restriction sites from adjacent cDNA fragments, resulting in an intact full-length cDNA of ZIKV. RNA transcripts derived from the full-length cDNA were infectious. The recombinant virus replicated as efficiently as the wild-type virus with similar growth kinetics and plaque morphologies in Vero and C6/36 cells. Both viruses were inhibited by NITD008 treatment. This in vitro ligation method will facilitate manipulation of the viral genome through genetic modifications of four separated subclones of ZIKV for the rapid and rational development of candidate vaccines and viral replication study.

  10. Hide and seek: a comparative autoradiographic in vitro investigation of the adenosine A3 receptor

    Energy Technology Data Exchange (ETDEWEB)

    Haeusler, D.; Fuchshuber, F.; Girschele, F.; Hacker, M.; Wadsak, W.; Mitterhauser, Markus [Medical University of Vienna, Department of Biomedical Imaging and Image-guided Therapy, Vienna (Austria); Grassinger, L. [University of Applied Sciences Wiener Neustadt, Department of Biomedical Analytics, Wiener Neustadt (Austria); Hoerleinsberger, W.J. [Medical University of Vienna, Department of Biomedical Imaging and Image-guided Therapy, Vienna (Austria); University of Vienna, Cognitive Science Research Platform, Vienna (Austria); Hoeftberger, R.; Leisser, I. [Medical University of Vienna, Institute of Neurology, Vienna (Austria); Shanab, K.; Spreitzer, H. [University of Vienna, Department of Drug and Natural Product Synthesis, Vienna (Austria); Gerdenitsch, W. [Medical University of Vienna, Institute of Biomedicinal Research, Vienna (Austria)

    2015-05-01

    Since the adenosine A3 receptor (A3R) is considered to be of high clinical importance in the diagnosis and treatment of ischaemic conditions (heart and brain), glaucoma, asthma, arthritis, cancer and inflammation, a suitable and selective A3R PET tracer such as [{sup 18}F]FE rate at SUPPY would be of high clinical value for clinicians as well as patients. A3R was discovered in the late 1990s, but there is still little known regarding its distribution in the CNS and periphery. Hence, in autoradiographic experiments the distribution of A3R in human brain and rat tissues was investigated and the specific binding of the A3R antagonist FE rate at SUPPY and MRS1523 compared. Immunohistochemical staining (IHC) experiments were also performed to validate the autoradiographic findings. For autoradiographic competition experiments human post-mortem brain and rat tissues were incubated with [{sup 125}I]AB-MECA and highly selective compounds to block the other adenosine receptor subtypes. Additionally, IHC was performed with an A3 antibody. Specific A3R binding of MRS1523 and FE rate at SUPPY was found in all rat peripheral tissues examined with the highest amounts in the spleen (44.0 % and 46.4 %), lung (44.5 % and 45.0 %), heart (39.9 % and 42.9 %) and testes (27.4 % and 29.5 %, respectively). Low amounts of A3R were found in rat brain tissues (5.9 % and 5.6 %, respectively) and human brain tissues (thalamus 8.0 % and 9.1 %, putamen 7.8 % and 8.2 %, cerebellum 6.0 % and 7.8 %, hippocampus 5.7 % and 5.6 %, caudate nucleus 4.9 % and 6.4 %, cortex 4.9 % and 6.3 %, respectively). The outcome of the A3 antibody staining experiments complemented the results of the autoradiographic experiments. The presence of A3R protein was verified in central and peripheral tissues by autoradiography and IHC. The specificity and selectivity of FE rate at SUPPY was confirmed by direct comparison with MRS1523, providing further evidence that [{sup 18}F]FE rate at SUPPY may be a suitable A3 PET

  11. In vitro-in vivo-in silico approach in biopharmaceutical characterization of ibuprofen IR and SR tablets.

    Science.gov (United States)

    Beloica, Sofija; Cvijić, Sandra; Bogataj, Marija; Parojčić, Jelena

    2015-07-30

    Within the last decades, physiologically based pharmacokinetic models have emerged into a biopharmaceutical toolkit that has been proven useful in understanding how physicochemical, formulation and physiological factors affect oral drug absorption. The purpose of this study was to develop a drug specific physiologically based pharmacokinetic model that will allow mechanistic interpretation of oral absorption from dosage forms exhibiting different in vitro and different in vivo performance (i.e. immediate release and sustained release tablets) and identification of bioperformance dissolution testing. Ibuprofen was chosen to be used for the "proof of concept" considering it is well characterised and the necessary physicochemical, biopharmaceutical and pharmacokinetic properties for model development could be found in the literature. Gastrointestinal simulation technology implemented in Simcyp® was successful in estimating ibuprofen oral absorption. The developed model exhibited good generalisation ability for the dosage forms studied. The obtained results indicate that the model was sensitive to input kinetics represented by the in vitro drug release profiles obtained under various dissolution conditions. According to the obtained results, reciprocating cylinder apparatus with biorepresentative change in media pH might be considered as bioperformance dissolution in the case of the two ibuprofen SR products studied. These results further justify the use of integrated in vitro-in vivo-in silico approach in estimating bioperformance of oral solid dosage forms. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Evaluation of Nystatin Containing Chitosan Hydrogels as Potential Dual Action Bio-Active Restorative Materials: in Vitro Approach

    Directory of Open Access Journals (Sweden)

    V. Tamara Perchyonok

    2014-11-01

    Full Text Available Healing is a specific biological process related to the general phenomenon of growth and tissue regeneration and is a process generally affected by several systemic conditions or as detrimental side-effects of chemotherapy- and radiotherapy-induced inflammation of the oral mucosa. The objectives of this study is to evaluate the novel chitosan based functional drug delivery systems, which can be successfully incorporated into “dual action bioactive restorative materials”, capable of inducing in vitro improved wound healing prototype and containing an antibiotic, such as nystatin, krill oil as an antioxidant and hydroxyapatite as a molecular bone scaffold, which is naturally present in bone and is reported to be successfully used in promoting bone integration when implanted as well as promoting healing. The hydrogels were prepared using a protocol as previously reported by us. The physico-chemical features, including surface morphology (SEM, release behaviors, stability of the therapeutic agent-antioxidant-chitosan, were measured and compared to the earlier reported chitosan-antioxidant containing hydrogels. Structural investigations of the reactive surface of the hydrogel are reported. Release of nystatin was investigated for all newly prepared hydrogels. Bio-adhesive studies were performed in order to assess the suitability of these designer materials. Free radical defense capacity of the biomaterials was evaluated using established in vitro model. The bio-adhesive capacity of the materials in the in vitro system was tested and quantified. It was found that the favorable synergistic effect of free radical built-in defense mechanism of the new functional materials increased sustainable bio-adhesion and therefore acted as a functional multi-dimensional restorative material with potential application in wound healing in vitro.

  13. Design and Investigation of PolyFermS In Vitro Continuous Fermentation Models Inoculated with Immobilized Fecal Microbiota Mimicking the Elderly Colon.

    Directory of Open Access Journals (Sweden)

    Sophie Fehlbaum

    Full Text Available In vitro gut modeling is a useful approach to investigate some factors and mechanisms of the gut microbiota independent of the effects of the host. This study tested the use of immobilized fecal microbiota to develop different designs of continuous colonic fermentation models mimicking elderly gut fermentation. Model 1 was a three-stage fermentation mimicking the proximal, transverse and distal colon. Models 2 and 3 were based on the new PolyFermS platform composed of an inoculum reactor seeded with immobilized fecal microbiota and used to continuously inoculate with the same microbiota different second-stage reactors mounted in parallel. The main gut bacterial groups, microbial diversity and metabolite production were monitored in effluents of all reactors using quantitative PCR, 16S rRNA gene 454-pyrosequencing, and HPLC, respectively. In all models, a diverse microbiota resembling the one tested in donor's fecal sample was established. Metabolic stability in inoculum reactors seeded with immobilized fecal microbiota was shown for operation times of up to 80 days. A high microbial and metabolic reproducibility was demonstrated for downstream control and experimental reactors of a PolyFermS model. The PolyFermS models tested here are particularly suited to investigate the effects of environmental factors, such as diet and drugs, in a controlled setting with the same microbiota source.

  14. Taguchi approach for anti-heat stress prescription compatibility in mice spleen lymphocytes in vitro.

    Science.gov (United States)

    Zhu, Xiao-Yu; Cheng, Gui-Lin; Liu, Feng-Hua; Yu, Jin; Wang, Yu-Jie; Yu, Tong-Quan; Xu, Jian-Qin; Wang, Ming

    2011-07-01

    Heat stress (HS) may induce immunosuppression as well as inhibit the proliferation of lymphocytes. This study evaluated the effects on immune function of our prescription on splenic lymphocytes under HS as well as its compatibility. The effects of four herbal extracts from Agastache rugosa, Atractylodes lancea, Cortex Phellodendri, and Gypsum Fibrosum on heat treated splenic lymphocytes were investigated and the compatibility of the prescription was also explored by using the Taguchi method. This study revealed changes in proliferation by traditional Chinese medicines of splenic lymphocytes after HS. Proliferation in the HS group was significantly lower than the control group. Under HS, the effects of higher concentrations of Agastache rugosa (100 and 200 μg/mL), Atractylodes lancea (100 and 200 μg/mL), Cortex Phellodendri (50 and 100 μg/mL) and Gypsum Fibrosum (100 and 200 μg/mL) caused a significant increase on ConA/LPS-induced proliferation of lymphocytes than lower concentrations. We, therefore, conclude that the prescription of traditional Chinese medicines may recover splenic lymphocytes from the immunosuppression induced by HS. The Taguchi design, which allows rapid and high efficiency for the selection of the best conditions for our prescription on HS-treated splenic lymphocytes, demonstrated that Agastache rugosa (200 μg/mL), Atractylodes lancea (200 μg/mL), Cortex Phellodendri (100 μg/mL) and Gypsum Fibrosum (100 μg/mL) were the optimal conditions for the prescription. The validation experiment confirmed that our composition in optimum extraction conditions enhanced effects on ConA or LPS-stimulated lymphocytes under HS. The results showed that the Taguchi optimization approach is a suitable method for optimization of the composition of prescription.

  15. Magnetic core/shell nanoparticle thin films deposited by MAPLE: Investigation by chemical, morphological and in vitro biological assays

    Energy Technology Data Exchange (ETDEWEB)

    Cristescu, R., E-mail: rodica.cristescu@inflpr.ro [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Popescu, C.; Socol, G.; Iordache, I.; Mihailescu, I.N. [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Mihaiescu, D.E.; Grumezescu, A.M. [Faculty of Applied Chemistry and Materials Science, ' Politehnica' University of Bucharest, 1-7 Polizu Street, 011061 Bucharest (Romania); Balan, A.; Stamatin, I. [University of Bucharest, 3Nano-SAE Research Center, PO Box MG-38, Bucharest-Magurele (Romania); Chifiriuc, C. [Faculty of Biology, University of Bucharest, Microbiology Immunology Department, Aleea Portocalilor 1-3, Sector 5, 77206 Bucharest (Romania); Bleotu, C. [Stefan S. Nicolau Institute of Virology, 285 Mihai Bravu, 030304 Bucharest (Romania); Saviuc, C.; Popa, M. [Faculty of Biology, University of Bucharest, Microbiology Immunology Department, Aleea Portocalilor 1-3, Sector 5, 77206 Bucharest (Romania); Chrisey, D.B. [Rensselaer Polytechnic Institute, School of Engineering, Departments of Materials Science and Biomedical Engineering, Troy, 12180-3590, NY (United States)

    2012-09-15

    Highlights: Black-Right-Pointing-Pointer We deposit magnetic Fe{sub 3}O{sub 4}/oleic acid/cephalosporin nanoparticle thin films by MAPLE. Black-Right-Pointing-Pointer Thin films have a chemical structure similar to the starting material. Black-Right-Pointing-Pointer Cephalosporins have an additive effect on the grain size and induce changes in grain shape. Black-Right-Pointing-Pointer MAPLE can be used to develop novel strategies for fighting medical biofilms associated with chronic infections. - Abstract: We report on thin film deposition of nanostructured Fe{sub 3}O{sub 4}/oleic acid/ceftriaxone and Fe{sub 3}O{sub 4}/oleic acid/cefepime nanoparticles (core/shell/adsorption-shell) were fabricated by matrix assisted pulsed laser evaporation (MAPLE) onto inert substrates. The thin films were characterized by profilometry, Fourier transform infrared spectroscopy, atomic force microscopy, and investigated by in vitro biological assays. The biological properties tested included the investigation of the microbial viability and the microbial adherence to the glass coverslip nanoparticle film, using Gram-negative and Gram-positive bacterial strains with known antibiotic susceptibility behavior, the microbial adherence to the HeLa cells monolayer grown on the nanoparticle pellicle, and the cytotoxicity on eukaryotic cells. The proposed system, based on MAPLE, could be used for the development of novel anti-microbial materials or strategies for fighting pathogenic biofilms frequently implicated in the etiology of biofilm associated chronic infections.

  16. Investigation of Patient-Specific Cerebral Aneurysm using Volumetric PIV, CFD, and In Vitro PC-MRI

    Science.gov (United States)

    Brindise, Melissa; Dickerhoff, Ben; Saloner, David; Rayz, Vitaliy; Vlachos, Pavlos

    2017-11-01

    4D PC-MRI is a modality capable of providing time-resolved velocity fields in cerebral aneurysms in vivo. The MRI-measured velocities and subsequent hemodynamic parameters such as wall shear stress, and oscillatory shear index, can help neurosurgeons decide a course of treatment for a patient, e.g. whether to treat or monitor the aneurysm. However, low spatiotemporal resolution, limited velocity dynamic range, and inherent noise of PC-MRI velocity fields can have a notable effect on subsequent calculations, and should be investigated. In this work, we compare velocity fields obtained with 4D PC-MRI, computational fluid dynamics (CFD) and volumetric particle image velocimetry (PIV), using a patient-specific model of a basilar tip aneurysm. The same in vitro model is used for all three modalities and flow input parameters are controlled. In vivo, PC-MRI data was also acquired for this patient and used for comparison. Specifically, we investigate differences in the resulting velocity fields and biases in subsequent calculations. Further, we explore the effect these errors may have on assessment of the aneurysm progression and seek to develop corrective algorithms and other methodologies that can be used to improve the accuracy of hemodynamic analysis in clinical setting.

  17. In vitro investigation of enhanced hemocompatibility and endothelial cell proliferation associated with quinone-rich polydopamine coating.

    Science.gov (United States)

    Luo, Rifang; Tang, Linlin; Zhong, Si; Yang, Zhilu; Wang, Jin; Weng, Yajun; Tu, Qiufen; Jiang, Chongxi; Huang, Nan

    2013-03-13

    Recent investigations have demonstrated that polydopamine (PDA)-modified surfaces were beneficial to the proliferation of endothelial cells (ECs). In this work, PDA coated 316L stainless steels (316L SS) were thermally treated at 50, 100, and 150 °C respectively (hereafter designated as Th50, Th100, and Th150) and consequently produced diverse surface chemical components. In vitro hemocompatibility and vascular cell-material interactions with ECs and smooth muscle cells (SMCs) affected by surface characteristics have been investigated. The Th150, rich in quinone, showed the best hemocompatibility and could effectively inhibit platelet adhesion, activation, and fibrinogen conformation transition. The polydopamine-modified surfaces were found to induce dramatic cell-material interaction with enhanced ECs proliferation, viability and migration, release of nitric oxide (NO), and reduced SMCs proliferation. The inhibitory effect of SMCs proliferation might be associated with the surface catechol content. The coating on Th150 showed a good resistance to the deformation of compression and expansion of vascular stents. These results effectively suggested that the Th150 coating might be promising when served as a stent coating platform.

  18. In vitro to in vivo extrapolation of effective dosimetry in developmental toxicity testing: Application of a generic PBK modelling approach.

    Science.gov (United States)

    Fragki, Styliani; Piersma, Aldert H; Rorije, Emiel; Zeilmaker, Marco J

    2017-10-01

    Incorporation of kinetics to quantitative in vitro to in vivo extrapolations (QIVIVE) is a key step for the realization of a non-animal testing paradigm, in the sphere of regulatory toxicology. The use of Physiologically-Based Kinetic (PBK) modelling for determining systemic doses of chemicals at the target site is accepted to be an indispensable element for such purposes. Nonetheless, PBK models are usually designed for a single or a group of compounds and are considered demanding, with respect to experimental data needed for model parameterization. Alternatively, we evaluate here the use of a more generic approach, i.e. the so-called IndusChemFate model, which is based on incorporated QSAR model parametrization. The model was used to simulate the in vivo kinetics of three diverse classes of developmental toxicants: triazoles, glycol ethers' alkoxyacetic acid metabolites and phthalate primary metabolites. The model required specific input per each class of compounds. These compounds were previously tested in three alternative assays: the whole-embryo culture (WEC), the zebrafish embryo test (ZET), and the mouse embryonic stem cell test (EST). Thereafter, the PBK-simulated blood levels at toxic in vivo doses were compared to the respective in vitro effective concentrations. Comparisons pertaining to relative potency and potency ranking with integration of kinetics were similar to previously obtained comparisons. Additionally, all three in vitro systems produced quite comparable results, and hence, a combination of alternative tests is still preferable for predicting the endpoint of developmental toxicity in vivo. This approach is put forward as biologically more plausible since plasma concentrations, rather than external administered doses, constitute the most direct in vivo dose metric. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Investigating the role of retinal Müller cells with approaches in genetics and cell biology.

    Science.gov (United States)

    Fu, Suhua; Zhu, Meili; Ash, John D; Wang, Yunchang; Le, Yun-Zheng

    2014-01-01

    Müller cells are major macroglia and play many essential roles as a supporting cell in the retina. As Müller cells only constitute a small portion of retinal cells, investigating the role of Müller glia in retinal biology and diseases is particularly challenging. To overcome this problem, we first generated a Cre/lox-based conditional gene targeting system that permits the genetic manipulation and functional dissection of gene of interests in Müller cells. To investigate diabetes-induced alteration of Müller cells, we recently adopted methods to analyze Müller cells survival/death in vitro and in vivo. We also used normal and genetically altered primary cell cultures to reveal the mechanistic insights for Müller cells in biological and disease processes. In this article, we will discuss the applications and limitations of these methodologies, which may be useful for research in retinal Müller cell biology and pathophysiology.

  20. Non-Invasive Assessment of Skin Barrier Properties: Investigating Emerging Tools for In Vitro and In Vivo Applications

    Directory of Open Access Journals (Sweden)

    Emer Duffy

    2017-10-01

    Full Text Available There is increasing interest in the development of non-invasive tools for studying the properties of skin, due to the potential for non-destructive sampling, reduced ethical concerns and the potential comparability of results in vivo and in vitro. The present research focuses on the use of a range of non-invasive approaches for studying skin and skin barrier properties in human skin and human skin equivalents (HSE. Analytical methods used include pH measurements, electrical sensing of the epidermis and detection of volatile metabolic skin products. Standard probe based measurements of pH and the tissue dielectric constant (TDC are used. Two other more novel approaches that utilise wearable platforms are also demonstrated here that can assess the electrical properties of skin and to profile skin volatile species. The potential utility of these wearable tools that permit repeatability of testing and comparability of results is considered through application of our recently reported impedance-based tattoo sensors and volatile samplers on both human participants and HSEs. The HSE exhibited a higher pH (6.5 and TDC (56 than human skin (pH 4.9–5.6, TDC 29–36, and the tattoo sensor revealed a lower impedance signal for HSEs, suggesting the model could maintain homeostasis, but in a different manner to human skin, which demonstrated a more highly resistive barrier. Characterisation of volatiles showed a variety of compound classes emanating from skin, with 16 and 27 compounds identified in HSEs and participants respectively. The continuing development of these tools offers potential for improved quality and relevance of data, and potential for detection of changes that are undetectable in traditional palpable and visual assessments, permitting early detection of irritant reactions.

  1. Multivariate approach to assess in vitro Fe bioaccessibility in chicken meat

    Directory of Open Access Journals (Sweden)

    Náira da Silva CAMPOS

    Full Text Available Abstract A 32 factorial design was employed to develop an in vitro digestion method for estimation of Fe bioaccessible fractions in cooked chicken meat. The effects of sample size and the in vitro bioaccessible fractions of this essential element were evaluated. A sample preparation method employing a microwave assisted digestion with dilute nitric acid was used prior to total Fe determination. For the bioacessibility studies, the optimized procedure employed 7.5 g of sample and 6% w/v of an acid pepsin solution. This procedure was applied to two kinds of chicken meat samples: breast and liver. Flame Atomic Absorption Spectrometry was used to determine total and bioaccessible (chyme or soluble portion levels of iron in the samples. With respect to total Fe content, the bioaccessible fractions of Fe found in these samples were around 23% and 56 %, for breast and chicken liver, respectively. The chicken liver sample showed the highest total (400 ± 10 mg kg-1 and bioaccessible Fe contents (223 ± 18 mg kg-1 and stands out as a good source of this micronutrient.

  2. Calcium fluoride nanoparticles induced suppression of Streptococcus mutans biofilm: an in vitro and in vivo approach.

    Science.gov (United States)

    Kulshrestha, Shatavari; Khan, Shakir; Hasan, Sadaf; Khan, M Ehtisham; Misba, Lama; Khan, Asad U

    2016-02-01

    Biofilm formation on the tooth surface is the root cause of dental caries and periodontal diseases. Streptococcus mutans is known to produce biofilm which is one of the primary causes of dental caries. Acid production and acid tolerance along with exopolysaccharide (EPS) formation are major virulence factors of S. mutans biofilm. In the current study, calcium fluoride nanoparticles (CaF2-NPs) were evaluated for their effect on the biofilm forming ability of S. mutans in vivo and in vitro. The in vitro studies revealed 89 % and 90 % reduction in biofilm formation and EPS production, respectively. Moreover, acid production and acid tolerance abilities of S. mutans were also reduced considerably in the presence of CaF2-NPs. Confocal laser scanning microscopy and transmission electron microscopy images were in accordance with the other results indicating inhibition of biofilm without affecting bacterial viability. The qRT-PCR gene expression analysis showed significant downregulation of various virulence genes (vicR, gtfC, ftf, spaP, comDE) associated with biofilm formation. Furthermore, CaF2-NPs were found to substantially decrease the caries in treated rat groups as compared to the untreated groups in in vivo studies. Scanning electron micrographs of rat's teeth further validated our results. These findings suggest that the CaF2-NPs may be used as a potential antibiofilm applicant against S. mutans and may be applied as a topical agent to reduce dental caries.

  3. Microscale In Vitro Assays for the Investigation of Neutral Red Retention and Ethoxyresorufin-O-Deethylase of Biofuels and Fossil Fuels.

    Directory of Open Access Journals (Sweden)

    Sebastian Heger

    Full Text Available Only few information on the potential toxic effectiveness of biofuels are available. Due to increasing worldwide demand for energy and fuels during the past decades, biofuels are considered as a promising alternative for fossil fuels in the transport sector. Hence, more information on their hazard potentials are required to understand the toxicological impact of biofuels on the environment. In the German Cluster of Excellence "Tailor-made Fuels from Biomass" design processes for economical, sustainable and environmentally friendly biofuels are investigated. In an unique and interdisciplinary approach, ecotoxicological methods are applied to gain information on potential adverse environmental effects of biofuels at an early phase of their development. In the present study, three potential biofuels, ethyl levulinate, 2-methyltetrahydrofuran and 2-methylfuran were tested. Furthermore, we investigated a fossil gasoline fuel, a fossil diesel fuel and an established biodiesel. Two in vitro bioassays, one for assessing cytotoxicity and one for aryl hydrocarbon receptor agonism, so called dioxin-like activity, as measured by Ethoxyresorufin-O-Deethylase, were applied using the permanent fish liver cell line RTL-W1 (Oncorhynchus mykiss. The special properties of these fuel samples required modifications of the test design. Points that had to be addressed were high substance volatility, material compatibility and low solubility. For testing of gasoline, diesel and biodiesel, water accommodated fractions and a passive dosing approach were tested to address the high hydrophobicity and low solubility of these complex mixtures. Further work has to focus on an improvement of the chemical analyses of the fuel samples to allow a better comparison of any effects of fossil fuels and biofuels.

  4. Microscale In Vitro Assays for the Investigation of Neutral Red Retention and Ethoxyresorufin-O-Deethylase of Biofuels and Fossil Fuels.

    Science.gov (United States)

    Heger, Sebastian; Bluhm, Kerstin; Brendt, Julia; Mayer, Philipp; Anders, Nico; Schäffer, Andreas; Seiler, Thomas-Benjamin; Hollert, Henner

    Only few information on the potential toxic effectiveness of biofuels are available. Due to increasing worldwide demand for energy and fuels during the past decades, biofuels are considered as a promising alternative for fossil fuels in the transport sector. Hence, more information on their hazard potentials are required to understand the toxicological impact of biofuels on the environment. In the German Cluster of Excellence "Tailor-made Fuels from Biomass" design processes for economical, sustainable and environmentally friendly biofuels are investigated. In an unique and interdisciplinary approach, ecotoxicological methods are applied to gain information on potential adverse environmental effects of biofuels at an early phase of their development. In the present study, three potential biofuels, ethyl levulinate, 2-methyltetrahydrofuran and 2-methylfuran were tested. Furthermore, we investigated a fossil gasoline fuel, a fossil diesel fuel and an established biodiesel. Two in vitro bioassays, one for assessing cytotoxicity and one for aryl hydrocarbon receptor agonism, so called dioxin-like activity, as measured by Ethoxyresorufin-O-Deethylase, were applied using the permanent fish liver cell line RTL-W1 (Oncorhynchus mykiss). The special properties of these fuel samples required modifications of the test design. Points that had to be addressed were high substance volatility, material compatibility and low solubility. For testing of gasoline, diesel and biodiesel, water accommodated fractions and a passive dosing approach were tested to address the high hydrophobicity and low solubility of these complex mixtures. Further work has to focus on an improvement of the chemical analyses of the fuel samples to allow a better comparison of any effects of fossil fuels and biofuels.

  5. A CDT-Based Heuristic Zone Design Approach for Economic Census Investigators

    Directory of Open Access Journals (Sweden)

    Changixu Cheng

    2015-01-01

    Full Text Available This paper addresses a special zone design problem for economic census investigators that is motivated by a real-world application. This paper presented a heuristic multikernel growth approach via Constrained Delaunay Triangulation (CDT. This approach not only solved the barriers problem but also dealt with the polygon data in zoning procedure. In addition, it uses a new heuristic method to speed up the zoning process greatly on the premise of the required quality of zoning. At last, two special instances for economic census were performed, highlighting the performance of this approach.

  6. Microfluidics approach to investigate the role of dynamic similitude in osteocyte mechanobiology.

    Science.gov (United States)

    Middleton, Kevin; Kondiboyina, Avinash; Borrett, Michael; Cui, Yi; Mei, Xueting; You, Lidan

    2017-10-13

    Fluid flow is an important regulator of cell function and metabolism in many tissues. Fluid shear stresses have been used to level the mechanical stimuli applied in vitro with what occurs in vivo. However, these experiments often lack dynamic similarity, which is necessary to ensure the validity of the model. For interstitial fluid flow, the major requirement for dynamic similarity is the Reynolds number (Re), the ratio of inertial to viscous forces, is the same between the system and model. To study the necessity of dynamic similarity for cell mechanotransduction studies, we investigated the response of osteocyte-like MLO-Y4 cells to different Re flows at the same level of fluid shear stress. Osteocytes were chosen for this study as flows applied in vitro and in vivo have Re that are orders of magnitude different. We hypothesize that osteocytes' response to fluid flow is Re dependent. We observed that cells exposed to lower and higher Re flows developed rounded and triangular morphologies, respectively. Lower Re flows also reduced apoptosis rates compared to higher Re flows. Furthermore, MLO-Y4 cells exposed to higher Re flows had stronger calcium responses compared to lower Re flows. However, by also controlling for flow rate, the lower Re flows induced a stronger calcium response; while degradation of components of the osteocyte glycocalyx reversed this effect. This work suggests that osteocytes are highly sensitive to differences in Re, independent of just shear stresses, supporting the need for improved in vitro flow platforms that better recapitulate the physiological environment. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  7. In vitro engineering of human 3D chondrosarcoma: a preclinical model relevant for investigations of radiation quality impact.

    Science.gov (United States)

    Hamdi, Dounia Houria; Barbieri, Sofia; Chevalier, François; Groetz, Jean-Emmanuel; Legendre, Florence; Demoor, Magali; Galera, Philippe; Lefaix, Jean-Louis; Saintigny, Yannick

    2015-08-08

    The benefit of better ballistic and higher efficiency of carbon ions for cancer treatment (hadron-therapy) is asserted since decades, especially for unresectable or resistant tumors like sarcomas. However, hadron-therapy with carbon ions stays underused and raises some concerns about potential side effects for patients. Chondrosarcoma is a cartilaginous tumor, chemo- and radiation-resistant, that lacks reference models for basic and pre-clinical studies in radiation-biology. Most studies about cellular effects of ionizing radiation, including hadrons, were performed under growth conditions dramatically different from human homeostasis. Tridimensional in vitro models are a fair alternative to animal models to approach tissue and tumors microenvironment. By using a collagen matrix, standardized culture conditions, physiological oxygen tension and a well defined chondrosarcoma cell line, we developed a pertinent in vitro 3D model for hadron-biology studies. Low- and high-Linear Energy Transfer (LET) ionizing radiations from GANIL facilities of ~1 keV/μm and 103 ± 4 keV/μm were used respectively, at 2 Gy single dose. The impact of radiation quality on chondrosarcoma cells cultivated in 3D was analyzed on cell death, cell proliferation and DNA repair. A fair distribution of chondrosarcoma cells was observed in the whole 3D scaffold. Moreover, LET distribution in depth, for ions, was calculated and found acceptable for radiation-biology studies using this kind of scaffold. No difference in cell toxicity was observed between low- and high-LET radiations but a higher rate of proliferation was displayed following high-LET irradiation. Furthermore, 3D models presented a higher and longer induction of H2AX phosphorylation after 2 Gy of high-LET compared to low-LET radiations. The presented results show the feasibility and usefulness of our 3D chondrosarcoma model in the study of the impact of radiation quality on cell fate. The observed changes in our tissue-like model

  8. A new approach for cerumenolytic treatment in children: In vivo and in vitro study.

    Science.gov (United States)

    Soy, F K; Ozbay, C; Kulduk, E; Dundar, R; Yazıcı, H; Sakarya, E U

    2015-07-01

    To demonstrate the effects of various cerumenolytic solutions in vivo and in vitro and to measure the change in pain following treatment. The study was done as a single-centre, prospective and double-blind study. Among 1243 paediatric patients with total or nearly total occlusive plug in 4 years period, those who accepted endoscopic ear examination and cleaning via aspiration after a follow-up period of at least 10 days following treatment were included in the study. Day of total TM visualisation was noted and removal co-efficient was calculated. The pre and post-treatment pain levels of the patients were assessed using analogue chromatic continuous scale (ACCS). In the in vitro part, cerumen samples collected at equal amounts from 20 patients were treated at 36-400°C in 6 different tubes with the same solutions and their dissolution degrees were assessed over a period of 5 days (Hour 6, Hour 12, Hour 48, Hour 72, Hour 92, Hour 120). Additionally, the degree of resolution in the tube treated with distilled water was considered to be the control reference. In the in vivo part of the study, total TM visualisation was observed in Group 1 at 50.2% (Day 3), in Group at 57.1%, in Group at 62.3%, in Group at 44.3% and in Group 5 at 73.5%. The group with the lowest removal co-efficient was Group 5 (removal co-efficient=1.623). In reference to the ACCS pain scores of the patients, the intra-group change pre-post treatment was found statistically significant for all groups (p=0.008; p=0.0222; p=0.005; p=0.026; p=0.018). After statistical analysis between the groups the difference between Group 5 and other groups was found statistically significant (p=0.002; p=0.026; p=0.044; p=0.034). In the in vitro part of the study, the best dissolution was observed in Group 2. In our study, the best cerumenolytic solutions were identified to be glycerine 10cc+3% hydrogen peroxide 10cc+10% sodium bicarbonate 10cc+distilled water 10cc. Especially the use of this mixture ease in terms of

  9. Size-dependent studies of macromolecular crowding on the thermodynamic stability, structure and functional activity of proteins: in vitro and in silico approaches.

    Science.gov (United States)

    Shahid, Sumra; Hassan, Md Imtaiyaz; Islam, Asimul; Ahmad, Faizan

    2017-02-01

    The environment inside cells in which proteins fold and function are quite different from that of the dilute buffer solutions often used during in vitro experiments. The presence of large amounts of macromolecules of varying shapes, sizes and compositions makes the intracellular milieu extremely crowded. The overall concentration of macromolecules ranges from 50 to 400gl-1, and they occupy 10-40% of the total cellular volume. These differences in solvent conditions and the level of crowdedness resulting in excluded volume effects can have significant consequences on proteins' biophysical properties. A question that arises is: how important is it to examine the roles of shape, size and composition of macromolecular crowders in altering the biological properties of proteins? This review article aims at focusing, gathering and summarizing all of the research investigations done by means of in vitro and in silico approaches taking into account the size-dependent influence of the crowders on proteins' properties. Altogether, the internal architecture of macromolecular crowding environment including size, shape and concentration of crowders, appears to be playing an extremely important role in causing changes in the biological processes. Most often the small sized crowders have been found more effective crowding agents. However, thermodynamic stability, structure and functional activity of proteins have been governed by volume exclusion as well as soft (chemical) interactions. The article provides an understanding of importance of internal architecture of the cellular environment in altering the biophysical properties of proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Investigating the Effects of Loading Factors on the In Vitro Pharmaceutical Performance of Mesoporous Materials as Drug Carriers for Ibuprofen

    Directory of Open Access Journals (Sweden)

    Junmin Lai

    2017-02-01

    Full Text Available The aim of the study was to investigate the effects of the loading factors, i.e., the initial drug loading concentration and the ratio of the drug to carriers, on the in vitro pharmaceutical performance of drug-loaded mesoporous systems. Ibuprofen (IBU was used as a model drug, and two non-ordered mesoporous materials of commercial silica Syloid® 244FP (S244FP and Neusilin® US2 (NS2 were selected in the study. The IBU-loaded mesoporous samples were prepared by a solvent immersion method with a rotary evaporation drying technique and characterized by polarized light microscopy (PLM, Fourier transform infrared (FTIR spectroscopy, X-ray powder diffraction (XRPD and differential scanning calorimetry (DSC. Dissolution experiments were performed in simulated gastric media at 37 °C under non-sink conditions. The concentration of IBU in solution was determined by HPLC. The study showed that the dissolution rate of IBU can be improved significantly using the mesoporous S224FP carriers due to the conversion of crystalline IBU into the amorphous form. Both of the loading factors affected the IBU dissolution kinetics. Due to the molecular interaction between the IBU and NS2 carriers, the loading factors had little effects on the drug release kinetics with incomplete drug desorption recovery and insignificant dissolution enhancement. Care and extensive evaluation must therefore be taken when mesoporous materials are chosen as carrier delivery systems.

  11. Carbohydrate Derived Fulvic Acid: An in vitro Investigation of a Novel Membrane Active Antiseptic Agent Against Candida albicans Biofilms.

    Science.gov (United States)

    Sherry, Leighann; Jose, Anto; Murray, Colin; Williams, Craig; Jones, Brian; Millington, Owain; Bagg, Jeremy; Ramage, Gordon

    2012-01-01

    Carbohydrate derived fulvic acid (CHD-FA) is a heat stable low molecular weight, water soluble, cationic, colloidal material with proposed therapeutic properties. The aim of this study was to evaluate the antifungal activity of CHD-FA against Candida albicans, and to characterize its mode of action. A panel of C. albicans isolates (n = 50) derived from a range of clinical specimens were grown planktonically and as biofilms, and the minimum inhibitory concentrations determined. Scanning electron microscopy was performed to examine ultrastructural changes and different cell membrane assays were used to determine its mode of action. In addition, the role of C. albicans biofilm resistance mechanisms were investigated to determine their effects on CHD-FA activity. CHD-FA was active against planktonic and sessile C. albicans at concentrations 0.125 and 0.25% respectively, and was shown to be fungicidal, acting through disruption of the cell membrane activity. Resistance mechanisms, including matrix, efflux, and stress, had a limited role upon CHD-FA activity. Overall, based on the promising in vitro spectrum of activity and minimal biofilm resistance of the natural and cheap antiseptic CHD-FA, further studies are required to determine its applicability for clinical use.

  12. In vitro investigation of molecules involved in Lactobacillus gasseri SBT2055 adhesion to host intestinal tract components.

    Science.gov (United States)

    Arai, T; Obuchi, S; Eguchi, K; Seto, Y

    2016-06-01

    The adhesion ability of Lactobacillus gasseri SBT2055 was investigated in vitro by searching for its adhesion molecules. Lactobacillus gasseri SBT2055 showed adherence to host components, including two commercially available mucins, Caco-2 epithelial-like cells and the extracellular matrix molecule fibronectin (Fn). Its adhesion rates to host components were generally higher than those of other Lactobacillus strains. We examined sortase-dependent proteins (SDPs) anchored by a sortase enzyme encoded by srtA1. The adhesion rates of an srtA1 disruptant were lower than those of Lact. gasseri SBT2055, and the relative adherences were as follows: two mucins, 43 and 40%; Caco-2, 66% and Fn, 28%. Seven additional gene disruptants were generated to determine the precise SDPs that contribute to adhesion to each component. The adhesion ability of Lact. gasseri SBT2055 was superior to those of other Lactobacillus strains. Additionally, four adhesion molecules were newly identified from candidate SDPs. Although the contribution of SDPs to adhesion has been reported using sortase gene disruptants, this is the first report to identify the precise SDPs that act as adhesion molecules. Our results will contribute to achieving better understanding of probiotic bacterial adherence. © 2016 The Society for Applied Microbiology.

  13. A New Approach for On-Demand Generation of Various Oxygen Tensions for In Vitro Hypoxia Models.

    Science.gov (United States)

    Li, Chunyan; Chaung, Wayne; Mozayan, Cameron; Chabra, Ranjeev; Wang, Ping; Narayan, Raj K

    2016-01-01

    The development of in vitro disease models closely mimicking the functions of human disease has captured increasing attention in recent years. Oxygen tensions and gradients play essential roles in modulating biological systems in both physiologic and pathologic events. Thus, controlling oxygen tension is critical for mimicking physiologically relevant in vivo environments for cell, tissue and organ research. We present a new approach for on-demand generation of various oxygen tensions for in vitro hypoxia models. Proof-of-concept prototypes have been developed for conventional cell culture microplate by immobilizing a novel oxygen-consuming biomaterial on the 3D-printed insert. For the first time, rapid (~3.8 minutes to reach 0.5% O2 from 20.9% O2) and precisely controlled oxygen tensions/gradients (2.68 mmHg per 50 μm distance) were generated by exposing the biocompatible biomaterial to the different depth of cell culture media. In addition, changing the position of 3D-printed inserts with immobilized biomaterials relative to the cultured cells resulted in controllable and rapid changes in oxygen tensions (approach allows enhanced spatiotemporal resolution and accuracy of the oxygen tensions. Additionally, it does not interfere with the testing environment while maintaining ease of use. The elegance of oxygen tension manipulation introduced by our new approach will drastically improve control and lower the technological barrier of entry for hypoxia studies. Since the biomaterials can be immobilized in any devices, including microfluidic devices and 3D-printed tissues or organs, it will serve as the basis for a new generation of experimental models previously impossible or very difficult to implement.

  14. Investigating the Joint Development of Approach Bias and Adolescent Alcohol Use.

    Science.gov (United States)

    Janssen, Tim; Wood, Mark D; Larsen, Helle; Peeters, Margot; Vollebergh, Wilma A M; Wiers, Reinout W

    2015-12-01

    We investigated the joint development between implicit approach bias and early adolescent alcohol use, and examined whether the link between approach bias and alcohol use was moderated by working memory (WM). The current study used data from a 2-year, 4-wave online sample of 378 Dutch early adolescents (mean age 14.9 years, 64.8% female). First, using latent growth curve modeling, we examined trajectories of approach bias and alcohol use over time. Second, we examined relations between baseline approach bias and WM and the development of alcohol use. Third, we examined the joint development of approach bias and alcohol use. Fourth, we examined whether the nature of this joint development varied for different levels of WM. Unconditional growth curve model analyses indicated that the functional forms of alcohol use and cognitive bias were best captured by quadratic and linear trajectories, respectively. We found that cognitive bias decreased over time. We found no significant relations between baseline predictors and observed increases in alcohol use. We found relations between the intercepts, but not to growth factors, in the joint development of alcohol use and approach bias. WM was not found to moderate relations between growth in approach bias and alcohol use in this sample. While we observed evidence of association between approach bias and alcohol use at baseline, there was no evidence of relations between development trajectories of the two. These findings replicate prior research demonstrating a role of implicit approach bias in predicting early adolescent alcohol use but do not demonstrate, in a light drinking early adolescent sample, the importance of interrelations between changes in approach bias and alcohol use over time, or a moderating role of WM. It is important to consider the potential consequences of repeated online approach bias assessment (e.g., changes in stimulus valence) when interpreting these results. Copyright © 2015 by the Research

  15. Combining in vitro and in silico approaches to evaluate the multifunctional profile of rosmarinic acid from Blechnum brasiliense on targets related to neurodegeneration.

    Science.gov (United States)

    Andrade, Juliana Maria de Mello; Dos Santos Passos, Carolina; Kieling Rubio, Maria Angélica; Mendonça, Jacqueline Nakau; Lopes, Norberto Peporine; Henriques, Amélia Teresinha

    2016-07-25

    Natural products are important sources of chemical diversity leading to unique scaffolds that can be exploited in the discovery of new drug candidates or chemical probes. In this context, chemical and biological investigation of ferns and lycophytes occurring in Brazil is an approach adopted by our research group aiming at discovering bioactive molecules acting on neurodegeneration targets. In the present study, rosmarinic acid (RA) isolated from Blechnum brasiliense showed an in vitro multifunctional profile characterized by antioxidant effects, and monoamine oxidases (MAO-A and MAO-B) and catechol-O-methyl transferase (COMT) inhibition. RA showed antioxidant effects against hydroxyl (HO(•)) and nitric oxide (NO) radicals (IC50 of 29.4 and 140 μM, respectively), and inhibition of lipid peroxidation (IC50 of 19.6 μM). In addition, RA inhibited MAO-A, MAO-B and COMT enzymes with IC50 values of 50.1, 184.6 and 26.7 μM, respectively. The MAO-A modulation showed a non-time-dependent profile, suggesting a reversible mechanism of inhibition. Structural insights on RA interactions with MAO-A and COMT were investigated by molecular docking. Finally, RA (up to 5 mM) demonstrated no cytotoxicity on polymorphonuclear rat cells. Taken together, our results suggest that RA may be exploited as a template for the development of new antioxidant molecules possessing additional MAO and COMT inhibition effects to be further investigated on in vitro and in vivo models of neurodegenerative diseases. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  16. Approaches to Learning at Work: Investigating Work Motivation, Perceived Workload, and Choice Independence

    Science.gov (United States)

    Kyndt, Eva; Raes, Elisabeth; Dochy, Filip; Janssens, Els

    2013-01-01

    Learning and development are taking up a central role in the human resource policies of organizations because of their crucial contribution to the competitiveness of those organizations. The present study investigates the relationship of work motivation, perceived workload, and choice independence with employees' approaches to learning at work.…

  17. Investigating the Efficiency of Scenario Based Learning and Reflective Learning Approaches in Teacher Education

    Science.gov (United States)

    Hursen, Cigdem; Fasli, Funda Gezer

    2017-01-01

    The main purpose of this research is to investigate the efficiency of scenario based learning and reflective learning approaches in teacher education. The impact of applications of scenario based learning and reflective learning on prospective teachers' academic achievement and views regarding application and professional self-competence…

  18. An Investigation of Interpersonal Problem Solving Approaches with Respect to Attachment Styles

    Science.gov (United States)

    Arslan, Emel; Arslan, Coskun; Ari, Ramazan

    2012-01-01

    The purpose of this study was to investigate interpersonal problem solving approaches with respect to attachment styles. The participants were 554 (279 female and 275 male) university students. Pearson product-moment correlation coefficients and multiple regression analysis were used. The results of the study showed that the attachment styles of…

  19. Performance Studies in Practice: An Investigation of Students' Approaches to Practice in a University Music Department

    Science.gov (United States)

    Burwell, Kim; Shipton, Matthew

    2011-01-01

    In higher education music, individual practice remains a unique and solitary activity, perhaps the least accessible to the scrutiny of either teachers or researchers. This study reports on an investigation of the variety among student approaches to practice, with reference to year groups, age, instrumental groups and examination results. Data were…

  20. An Investigation of Human-Computer Interaction Approaches Beneficial to Weak Learners in Complex Animation Learning

    Science.gov (United States)

    Yeh, Yu-Fang

    2016-01-01

    Animation is one of the useful contemporary educational technologies in teaching complex subjects. There is a growing interest in proper use of learner-technology interaction to promote learning quality for different groups of learner needs. The purpose of this study is to investigate if an interaction approach supports weak learners, who have…

  1. Student-Centred Learning Environments: An Investigation into Student Teachers' Instructional Preferences and Approaches to Learning

    Science.gov (United States)

    Baeten, Marlies; Dochy, Filip; Struyven, Katrien; Parmentier, Emmeline; Vanderbruggen, Anne

    2016-01-01

    The use of student-centred learning environments in education has increased. This study investigated student teachers' instructional preferences for these learning environments and how these preferences are related to their approaches to learning. Participants were professional Bachelor students in teacher education. Instructional preferences and…

  2. Investigating Acceptance toward Mobile Learning to Assist Individual Knowledge Management: Based on Activity Theory Approach

    Science.gov (United States)

    Liaw, Shu-Sheng; Hatala, Marek; Huang, Hsiu-Mei

    2010-01-01

    Mobile devices could facilitate human interaction and access to knowledge resources anytime and anywhere. With respect to wide application possibilities of mobile learning, investigating learners' acceptance towards it is an essential issue. Based on activity theory approach, this research explores positive factors for the acceptance of m-learning…

  3. A Visual Approach to Investigating Shared and Global Memory Behavior of CUDA Kernels

    KAUST Repository

    Rosen, Paul

    2013-06-01

    We present an approach to investigate the memory behavior of a parallel kernel executing on thousands of threads simultaneously within the CUDA architecture. Our top-down approach allows for quickly identifying any significant differences between the execution of the many blocks and warps. As interesting warps are identified, we allow further investigation of memory behavior by visualizing the shared memory bank conflicts and global memory coalescence, first with an overview of a single warp with many operations and, subsequently, with a detailed view of a single warp and a single operation. We demonstrate the strength of our approach in the context of a parallel matrix transpose kernel and a parallel 1D Haar Wavelet transform kernel. © 2013 The Author(s) Computer Graphics Forum © 2013 The Eurographics Association and Blackwell Publishing Ltd.

  4. Effects of sertraline and fluoxetine on p-glycoprotein at barrier sites: in vivo and in vitro approaches.

    Directory of Open Access Journals (Sweden)

    Amita Kapoor

    Full Text Available BACKGROUND AND PURPOSE: Retention of substances from systemic circulation in the brain and testes are limited due to high levels of P-glycoprotein (P-gp in the luminal membranes of brain and testes capillary endothelial cells. From a clinical perspective, P-gp rapidly extrudes lipophilic therapeutic agents, which then fail to reach efficacious levels. Recent studies have demonstrated that acute administration of selective serotonin reuptake inhibitors (SSRI can affect P-gp function, in vitro and in vivo. However, little is known concerning the time-course of these effects or the effects of different SSRI in vivo. EXPERIMENTAL APPROACH: The P-gp substrate, tritiated digoxin ([(3H] digoxin, was co-administered with fluoxetine or sertraline to determine if either compound increased drug accumulation within the brains and testes of mice due to inhibition of P-gp activity. We undertook parallel studies in endothelial cells derived from brain microvessels to determine the dose-response and time-course of effects. KEY RESULTS: In vitro, sertraline resulted in rapid and potent inhibition of P-gp function in brain endothelial cells, as determined by cellular calcein accumulation. In vivo, a biphasic effect was demonstrated. Brain accumulation of [(3H] digoxin was increased 5 minutes after treatment with sertraline, but by 60 minutes after sertraline treatment, brain accumulation of digoxin was reduced compared to control. By 240 minutes after sertraline treatment brain digoxin accumulation was elevated compared to control. A similar pattern of results was obtained in the testes. There was no significant effect of fluoxetine on P-gp function, in vitro or in vivo. CONCLUSIONS AND IMPLICATIONS: Acute sertraline administration can modulate P-gp activity in the blood-brain barrier and blood-testes barrier. This clearly has implications for the ability of therapeutic agents that are P-gp substrates, to enter the brain when co-administered with SSRI.

  5. Ultrasound effects on brain-targeting mannosylated liposomes: in vitro and blood–brain barrier transport investigations

    Directory of Open Access Journals (Sweden)

    Zidan AS

    2015-07-01

    Full Text Available Ahmed S Zidan,1,2 Hibah Aldawsari1 1Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia; 2Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt Abstract: Delivering drugs to intracerebral regions can be accomplished by improving the capacity of transport through blood–brain barrier. Using sertraline as model drug for brain targeting, the current study aimed at modifying its liposomal vesicles with mannopyranoside. Box-Behnken design was employed to statistically optimize the ultrasound parameters, namely ultrasound amplitude, time, and temperature, for maximum mannosylation capacity, sertraline entrapment, and surface charge while minimizing vesicular size. Moreover, in vitro blood–brain barrier transport model was established to assess the transendothelial capacity of the optimized mannosylated vesicles. Results showed a dependence of vesicular size, mannosylation capacity, and sertraline entrapment on cavitation and bubble implosion events that were related to ultrasound power amplitude, temperature. However, short ultrasound duration was required to achieve >90% mannosylation with nanosized vesicles (<200 nm of narrow size distribution. Optimized ultrasound parameters of 65°C, 27%, and 59 seconds for ultrasound temperature, amplitude, and time were elucidated to produce 81.1%, 46.6 nm, and 77.6% sertraline entrapment, vesicular size, and mannosylation capacity, respectively. Moreover, the transendothelial ability was significantly increased by 2.5-fold by mannosylation through binding with glucose transporters. Hence, mannosylated liposomes processed by ultrasound could be a promising approach for manufacturing and scale-up of brain-targeting liposomes. Keywords: CNS delivery, sizing, lipid based formulations, quality by design, sertraline hydrochloride

  6. An in vitro approach for modelling branchial copper binding in rainbow trout.

    Science.gov (United States)

    Taylor, Lisa N; Baker, Daniel W; Wood, Chris M; McDonald, D Gordon

    2002-09-01

    The main objective of this study was to characterize the individual effects of water chemistry (Ca(2+), Na(+), dissolved organic matter (DOM), pH, alkalinity) on the rapid binding of copper to the gill surface of rainbow trout using an in vitro gill binding assay. In this assay, individual gill arches were exposed for 5 min to (64)Cu labelled copper solutions ranging from 0.02 to 0.16 microM in water chemistries reflecting the full range of fresh water values for the Great Lakes. The gills displayed saturable Cu binding within this Cu range but gill-Cu binding was completely unaffected over the full range of calcium, sodium and alkalinity concentrations used. Only low pH (pH 4.0) and commercial DOM (Aldrich humic acid at > or =3 mgC/l) altered copper binding to rainbow trout gills in vitro. These findings were consistent with the results of geochemical modelling of our water chemistry (using MINEQL+, Version 4.5) which showed that H(+) and DOM affected the free cupric ion concentration. However, DOM (up to 80 mgC/l) was only able to reduce Cu on the gills by 50%. We hypothesize that in the range of 0.02-0.16 microM Cu there are two high affinity Cu binding sites on the gills, one having a substantially higher affinity for copper than DOM. The absence of a calcium effect on gill copper binding was in accord with in vivo evidence that calcium primarily acts to alter the physiology of the gill binding sites through acclimatory processes, rather than through competitive interactions. It was a surprise that water chemistry parameters influence rapid gill-metal binding in a manner different to their influence on acute toxicity and different from the effects on long-term binding reported in other studies. Currently, the biotic ligand model uses the rapid increase of gill copper (believed to reflect binding to the physiologically active receptor sites) to model gill binding characteristics. The distinction between rapid surface binding and metal uptake obviously plays an

  7. Inducers of salmon innate immunity: An in vitro and in vivo approach.

    Science.gov (United States)

    Estévez, Rosana A; Mostazo, Miriam G Contreras; Rodriguez, Eduardo; Espinoza, Juan Carlos; Kuznar, Juan; Jónsson, Zophonías O; Guðmundsson, Guðmundur H; Maier, Valerie H

    2018-01-01

    Maintaining fish health is one of the most important aims in aquaculture. Prevention of fish diseases therefore is crucial and can be achieved by various different strategies, including most often a combination of different methods such as optimal feed and fish density, as well as strengthening the immune system. Understanding the fish innate immune system and developing methods to activate it, in an effort to prevent infections in the first place, has been a goal in recent years. In this study we choose different inducers of the innate immune system and examined their effects in vitro on the salmon cell line CHSE-214. We found that the butyrate derivatives 4-phenyl butyrate (PBA) and β-hydroxy-β-methyl butyrate (HMB) induce the expression of various innate immune genes differentially over 24-72 h. Similarly, lipids generated from fish oils were found to have an effect on the expression of the antimicrobial peptides cathelicidin and hepcidin, as well as iNOS and the viral receptor RIG-1. Interestingly we found that vitamin D3, similar as in mammals, was able to increase cathelicidin expression in fish cells. The observed induction of these different innate immune factors correlated with antibacterial activity against Aeromonas salmonicida and antiviral activity against IPNV and ISAV in vitro. To relate this data to the in vivo situation we examined cathelicidin expression in juvenile salmon and found that salmon families vary greatly in their basal cathelicidin levels. Examining cathelicidin levels in families known to be resistant to IPNV showed that these QTL-families had lower basal levels of cathelicidin in gills, than non QTL-families. Feeding fish with HMB caused a robust increase in cathelicidin expression in gills, but not skin and this was independent of the fish being resistant to IPNV. These findings support the use of fish cell lines as a tool to develop new inducers of the fish innate immune system, but also highlight the importance of the tissue

  8. Safety profile of solid lipid nanoparticles loaded with rosmarinic acid for oral use: in vitro and animal approaches.

    Science.gov (United States)

    Madureira, Ana Raquel; Nunes, Sara; Campos, Débora A; Fernandes, João C; Marques, Cláudia; Zuzarte, Monica; Gullón, Beatriz; Rodríguez-Alcalá, Luís M; Calhau, Conceição; Sarmento, Bruno; Gomes, Ana Maria; Pintado, Maria Manuela; Reis, Flávio

    2016-01-01

    Rosmarinic acid (RA) possesses several protective bioactivities that have attracted increasing interest by nutraceutical/pharmaceutical industries. Considering the reduced bioavailability after oral use, effective (and safe) delivery systems are crucial to protect RA from gastrointestinal degradation. This study aims to characterize the safety profile of solid lipid nanoparticles produced with Witepsol and Carnauba waxes and loaded with RA, using in vitro and in vivo approaches, focused on genotoxicity and cytotoxicity assays, redox status markers, hematological and biochemical profile, liver and kidney function, gut bacterial microbiota, and fecal fatty acids composition. Free RA and sage extract, empty nanoparticles, or nanoparticles loaded with RA or sage extract (0.15 and 1.5 mg/mL) were evaluated for cell (lymphocytes) viability, necrosis and apoptosis, and antioxidant/prooxidant effects upon DNA. Wistar rats were orally treated for 14 days with vehicle (control) and with Witepsol or Carnauba nanoparticles loaded with RA at 1 and 10 mg/kg body weight/d. Blood, urine, feces, and several tissues were collected for analysis. Free and loaded RA, at 0.15 mg/mL, presented a safe profile, while genotoxic potential was found for the higher dose (1.5 mg/mL), mainly by necrosis. Our data suggest that both types of nanoparticles are safe when loaded with moderate concentrations of RA, without in vitro genotoxicity and cytotoxicity and with an in vivo safety profile in rats orally treated, thus opening new avenues for use in nutraceutical applications.

  9. Novel targeted approach to better understand how natural structural barriers govern carotenoid in vitro bioaccessibility in vegetable-based systems.

    Science.gov (United States)

    Palmero, Paola; Lemmens, Lien; Ribas-Agustí, Albert; Sosa, Carola; Met, Kristof; de Dieu Umutoni, Jean; Hendrickx, Marc; Van Loey, Ann

    2013-12-01

    An experimental approach, allowing us to understand the effect of natural structural barriers (cell walls, chromoplast substructures) on carotenoid bioaccessibility, was developed. Different fractions with different levels of carotenoid bio-encapsulation (carotenoid-enriched oil, chromoplasts, small cell clusters, and large cell clusters) were isolated from different types of carrots and tomatoes. An in vitro method was used to determine carotenoid bioaccessibility. In the present work, a significant decrease in carotenoid in vitro bioaccessibility could be observed with an increasing level of bio-encapsulation. Differences in cell wall material and chromoplast substructure between matrices influenced carotenoid release and inclusion in micelles. For carrots, cell walls and chromoplast substructure were important barriers for carotenoid bioaccessibility while, in tomatoes, the chromoplast substructure represented the most important barrier governing bioaccessibility. The highest increase in carotenoid bioaccessibility, for all matrices, was obtained after transferring carotenoids into the oil phase, a system lacking cell walls and chromoplast substructures that could hamper carotenoid release. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. In vitro functional assessment of natural HIV-1 group M Vpu sequences using a universal priming approach.

    Science.gov (United States)

    Rahimi, Asa; Anmole, Gursev; Soto-Nava, Maribel; Escamilla-Gomez, Tania; Markle, Tristan; Jin, Steven W; Lee, Guinevere Q; Harrigan, P Richard; Bangsberg, David R; Martin, Jeffrey; Avila-Rios, Santiago; Reyes-Teran, Gustavo; Brockman, Mark A; Brumme, Zabrina L

    2017-02-01

    The HIV-1 accessory protein Vpu exhibits high inter- and intra- subtype genetic diversity that may influence Vpu function and possibly contribute to HIV-1 pathogenesis. However, scalable methods to evaluate genotype/phenotype relationships in natural Vpu sequences are limited, particularly those expressing the protein in CD4+ T-cells, the natural target of HIV-1 infection. A major impediment to assay scalability is the extensive genetic diversity within, and immediately upstream of, Vpu's initial 5' coding region, which has necessitated the design of oligonucleotide primers specific for each individual HIV-1 isolate (or subtype). To address this, we developed two universal forward primers, located in relatively conserved regions 38 and 90 bases upstream of Vpu, and a single universal reverse primer downstream of Vpu, which are predicted to cover the vast majority of global HIV-1 group M sequence diversity. We show that inclusion of up to 90 upstream bases of HIV-1 genomic sequence does not significantly influence in vitro Vpu expression or function when a Rev/Rev Response Element (RRE)-dependent expression system is used. We further assess the function of four diverse HIV-1 Vpu sequences, revealing reproducible and significant differences between them. Our approach represents a scalable option to measure the in vitro function of genetically diverse natural Vpu isolates in a CD4+ T-cell line. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. "Cell Migration" Is the Ontology Group Differentially Expressed in Porcine Oocytes Before and After In Vitro Maturation: A Microarray Approach.

    Science.gov (United States)

    Kranc, Wiesława; Budna, Joanna; Chachuła, Adrian; Borys, Sylwia; Bryja, Artur; Rybska, Marta; Ciesiółka, Sylwia; Sumelka, Ewa; Jeseta, Michał; Brüssow, Klaus P; Bukowska, Dorota; Antosik, Paweł; Bruska, Małgorzata; Nowicki, Michał; Zabel, Maciej; Kempisty, Bartosz

    2017-04-01

    Maturation of cumulus-oocyte complexes (COCs) is crucial for further successful monospermic fertilization, embryo growth, and implantation. All these events are accompanied by proliferation and differentiation of cumulus cells. The migration of COCs to the oviduct after ovulation and the interaction between female gametes and/or embryos with maternal tissues are still poorly recognized on the molecular level. This study was aimed to first demonstrate the mRNA expression profile of cell migration markers during different stages of porcine oocytes maturation and developmental capability in vitro. The COCs were collected from a total of 45 pubertal crossbred Landrace gilts, brilliant cresyl blue (BCB) stained, and analyzed before (n = 150) or after (n = 150) in vitro maturation (IVM). Using the Affymetrix® Porcine Gene 1.1 ST Array, the expression profile of 12,258 porcine transcripts was examined. We found nine genes involved in cell migration mechanisms, that is, PLD1, KIT, LAMA2, MAP3K1, VEGFA, TGFBR3, INSR, TPM1, and RTN4. These genes were upregulated in porcine oocytes before IVM as compared with post-IVM expression analysis. Moreover, important mechanisms of biological interaction between VEGFA-KIT and VEGFA-INSR were also observed. The upregulation and/or downregulation of selected mRNAs expression after microarray assays was checked and approved by real-time quantitative polymerase chain reaction. We suggest that several genes, including LAMA2 or TPM1, encode proteins participating in the formation of the oocyte's protein architecture such as microtubules and kinetochore reorganization. As the expression of all "migration regulatory genes" investigated in this study was significantly upregulated in oocytes before IVM, we conclude that they may contribute to the maturational capability of porcine oocytes. However, migration potency of COCs is not accompanied by achievement of the MII stage by porcine oocytes in vitro. The investigated genes such as PLD

  12. Interaction of graphene-related materials with human intestinal cells: an in vitro approach.

    Science.gov (United States)

    Kucki, M; Rupper, P; Sarrieu, C; Melucci, M; Treossi, E; Schwarz, A; León, V; Kraegeloh, A; Flahaut, E; Vázquez, E; Palermo, V; Wick, P

    2016-04-28

    Graphene-related materials (GRM) inherit unique combinations of physicochemical properties which offer a high potential for technological as well as biomedical applications. It is not clear which physicochemical properties are the most relevant factors influencing the behavior of GRM in complex biological environments. In this study we have focused on the interaction of GRM, especially graphene oxide (GO), and Caco-2 cells in vitro. We mimiked stomach transition by acid-treatment of two representative GRM followed by analysis of their physicochemical properties. No significant changes in the material properties or cell viability of exposed Caco-2 cells in respect to untreated GRM could be detected. Furthermore, we explored the interaction of four different GO and Caco-2 cells to identify relevant physicochemical properties for the establishment of a material property-biological response relationship. Despite close interaction with the cell surface and the formation of reactive oxygen species (ROS), no acute toxicity was found for any of the applied GO (concentration range 0-80 μg ml(-1)) after 24 h and 48 h exposure. Graphene nanoplatelet aggregates led to low acute toxicity at high concentrations, indicating that aggregation, the number of layers or the C/O ratio have a more pronounced effect on the cell viability than the lateral size alone.

  13. Two-dimensional HPLC coupled to ICP-MS and electrospray ionisation (ESI)-MS/MS for investigating the bioavailability in vitro of arsenic species from edible seaweed

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Sartal, Cristina; Barciela-Alonso, Maria del Carmen; Bermejo-Barrera, Pilar [University of Santiago de Compostela, Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, Santiago de Compostela (Spain); Taebunpakul, Sutthinun [LGC Limited, Teddington, Middlesex (United Kingdom); Imperial College of Science, Technology and Medicine, South Kensington, Department of Materials, London (United Kingdom); National Institute of Metrology (Thailand), Pathumthani (Thailand); Stokes, Emma; Goenaga-Infante, Heidi [LGC Limited, Teddington, Middlesex (United Kingdom)

    2012-04-15

    Edible seaweed consumption is a route of exposure to arsenic. However, little attention has been paid to estimate the bioaccessibility and/or bioavailability of arsenosugars in edible seaweed and their possible degradation products during gastrointestinal digestion. This work presents first use of combined inductively coupled plasma mass spectroscopy (ICP-MS) with electrospray ionization tandem mass spectrometry (ESI-MS/MS) with two-dimensional HPLC (size exclusion followed by anion exchange) to compare the qualitative and quantitative arsenosugars speciation of different edible seaweed with that of their bioavailable fraction as obtained using an in vitro gastrointestinal digestion procedure. Optimal extraction conditions for As species from four seaweed namely kombu, wakame, nori and sea lettuce were selected as a compromise between As extraction efficiency and preservation of compound identity. For most investigated samples, the use of ammonium acetate buffer as extractant and 1 h sonication in a water bath followed by HPLC-ICP-MS resulted in 40-61% of the total As to be found in the buffered aqueous extract, of which 86-110% was present as arsenosugars (glycerol sugar, phosphate sugar and sulfonate sugar for wakame and kombu and glycerol sugar and phosphate sugar for nori). The exception was sea lettuce, for which the arsenosugar fraction (glycerol sugar, phosphate sugar) only comprised 44% of the total extracted As. Interestingly, the ratio of arsenobetaine and dimethylarsinic acid to arsenosugars in sea lettuce extracts seemed higher than that for the rest of investigated samples. After in vitro gastrointestinal digestion, approximately 11-16% of the total As in the solid sample was found in the dialyzates with arsenosugars comprising 93-120% and 41% of the dialyzable As fraction for kombu, wakame, nori and sea lettuce, respectively. Moreover, the relative As species distribution in seaweed-buffered extracts and dialyzates was found to be very similar

  14. Accuracy of Digital vs Conventional Implant Impression Approach: A Three-Dimensional Comparative In Vitro Analysis.

    Science.gov (United States)

    Basaki, Kinga; Alkumru, Hasan; De Souza, Grace; Finer, Yoav

    To assess the three-dimensional (3D) accuracy and clinical acceptability of implant definitive casts fabricated using a digital impression approach and to compare the results with those of a conventional impression method in a partially edentulous condition. A mandibular reference model was fabricated with implants in the first premolar and molar positions to simulate a patient with bilateral posterior edentulism. Ten implant-level impressions per method were made using either an intraoral scanner with scanning abutments for the digital approach or an open-tray technique and polyvinylsiloxane material for the conventional approach. 3D analysis and comparison of implant location on resultant definitive casts were performed using laser scanner and quality control software. The inter-implant distances and interimplant angulations for each implant pair were measured for the reference model and for each definitive cast (n = 20 per group); these measurements were compared to calculate the magnitude of error in 3D for each definitive cast. The influence of implant angulation on definitive cast accuracy was evaluated for both digital and conventional approaches. Statistical analysis was performed using t test (α = .05) for implant position and angulation. Clinical qualitative assessment of accuracy was done via the assessment of the passivity of a master verification stent for each implant pair, and significance was analyzed using chi-square test (α = .05). A 3D error of implant positioning was observed for the two impression techniques vs the reference model, with mean ± standard deviation (SD) error of 116 ± 94 μm and 56 ± 29 μm for the digital and conventional approaches, respectively (P = .01). In contrast, the inter-implant angulation errors were not significantly different between the two techniques (P = .83). Implant angulation did not have a significant influence on definitive cast accuracy within either technique (P = .64). The verification stent

  15. A single-tube approach for in vitro diagnostics using diatomaceous earth and optical sensor.

    Science.gov (United States)

    Zhao, Fei; Koo, Bonhan; Liu, Huifang; Eun Jin, Choong; Shin, Yong

    2018-01-15

    Versatile, simple and efficient sample preparation is desirable for point-of-care testing of emerging diseases such as zoonoses, but current sample preparation assays are insensitive, labour-intensive and time-consuming and require multiple instruments. We developed a single-tube sample preparation approach involving direct pathogen enrichment and extraction from human specimens using diatomaceous earth (DE). Amine-modified DE was used to directly enrich a zoonotic pathogen, Brucella, in a large sample volume. Next, a complex of amine-modified DE and dimethyl suberimidate was used for nucleic acid extraction from the enriched pathogen. Using our single-tube approach, the pathogen can be enriched and extracted within 60min at a level of 1 colony formation unit (CFU) from a 1ml sample volume in the same tube. The performance of this approach is 10-100 times better than that of a commercial kit (102 to 103CFU/ml) but does not require a large centrifuge. Finally, we combined the single-tube approach with a bio-optical sensor for rapid and accurate zoonotic pathogen detection in human urine samples. Using the combination system, Brucella in human urine can be efficiently enriched (~ 8-fold) and the detection limit is enhanced by up to 100 times (1CFU/ml bacteria in urine) compared with the commercial kit. This combined system is fast and highly sensitive and thus represents a promising approach for disease diagnosis in the clinical setting. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. New practical and theoretical approaches to the induction of morphogenesis from plant tumors in vitro using new types of plant growth regulators: towards constructive paradigms in agriculture and medicine.

    Science.gov (United States)

    Lieber, Michael M

    2013-01-01

    Using classical or traditional plant growth regulators, calli or plant tumors have been produced in vitro and subsequently have been induced to produce buds and plantlets, a process referred to as regeneration. For many years, this has been a successful procedure for in vitro, plant propagation. However, for a number of plant species investigators could not induce calli in vitro to produce buds. Organogenesis was still recalcitrant for various plants in 1980. New types or nonconventional growth regulators, such as methylglyoxal and ascorbic acid, were then found to overcome recalcitrant organogenesis in vitro. Their successful or effective use gave support to a theory that stressful, non-uniform cohesive force-fields, electromagnetic in nature, occurring through the application of certain chemicals, are necessary for in vitro morphogenesis from plant neoplasm or callus. Morphogenesis is seen as an adaptive accommodation to the inner stresses from such non-uniform, cohesive forces. Diverse chemicals, not considered traditional plant growth regulators would, it has been conjectured, enable the generation of such cohesive forces, in non-uniform arrays, and it has been predicted that more chemicals of this type will be discovered. A new constructive approach to agriculture and medicine, using a new plant tissue-culture model, based on new theory, has also been predicted.

  17. Management Approach for NASA's Earth Venture-1 (EV-1) Airborne Science Investigations

    Science.gov (United States)

    Guillory, Anthony R.; Denkins, Todd C.; Allen, B. Danette

    2013-01-01

    The Earth System Science Pathfinder (ESSP) Program Office (PO) is responsible for programmatic management of National Aeronautics and Space Administration's (NASA) Science Mission Directorate's (SMD) Earth Venture (EV) missions. EV is composed of both orbital and suborbital Earth science missions. The first of the Earth Venture missions is EV-1, which are Principal Investigator-led, temporally-sustained, suborbital (airborne) science investigations costcapped at $30M each over five years. Traditional orbital procedures, processes and standards used to manage previous ESSP missions, while effective, are disproportionally comprehensive for suborbital missions. Conversely, existing airborne practices are primarily intended for smaller, temporally shorter investigations, and traditionally managed directly by a program scientist as opposed to a program office such as ESSP. In 2010, ESSP crafted a management approach for the successful implementation of the EV-1 missions within the constructs of current governance models. NASA Research and Technology Program and Project Management Requirements form the foundation of the approach for EV-1. Additionally, requirements from other existing NASA Procedural Requirements (NPRs), systems engineering guidance and management handbooks were adapted to manage programmatic, technical, schedule, cost elements and risk. As the EV-1 missions are nearly at the end of their successful execution and project lifecycle and the submission deadline of the next mission proposals near, the ESSP PO is taking the lessons learned and updated the programmatic management approach for all future Earth Venture Suborbital (EVS) missions for an even more flexible and streamlined management approach.

  18. Molecular Architecture of the Blood Brain Barrier Tight Junction Proteins--A Synergistic Computational and In Vitro Approach.

    Science.gov (United States)

    Irudayanathan, Flaviyan Jerome; Trasatti, John P; Karande, Pankaj; Nangia, Shikha

    2016-01-14

    The blood-brain barrier (BBB) constituted by claudin-5 tight junctions is critical in maintaining the homeostasis of the central nervous system, but this highly selective molecular interface is an impediment for therapeutic interventions in neurodegenerative and neurological diseases. Therapeutic strategies that can exploit the paracellular transport remain elusive due to lack of molecular insights of the tight junction assembly. This study focuses on analyzing the membrane driven cis interactions of claudin-5 proteins in the formation of the BBB tight junctions. We have adopted a synergistic approach employing in silico multiscale dynamics and in vitro cross-linking experiments to study the claudin-5 interactions. Long time scale simulations of claudin-5 monomers, in seven different lipid compositions, show formation of cis dimers that subsequently aggregate into strands. In vitro formaldehyde cross-linking studies also conclusively show that cis-interacting claudin-5 dimers cross-link with short methylene spacers. Using this synergistic approach, we have identified five unique dimer interfaces in our simulations that correlate with the cross-linking experiments, four of which are mediated by transmembrane (TM) helices and the other mediated by extracellular loops (ECL). Potential of mean force calculations of these five dimers revealed that the TM mediated interfaces, which can have distinctive leucine zipper interactions in some cases, are more stable than the ECL mediated interface. Additionally, simulations show that claudin-5 dimerization is significantly influenced by the lipid microenvironment. This study captures the fundamental interactions responsible for the BBB tight junction assembly and offers a framework for extending this work to other tight junctions found in the body.

  19. "Bone Development" Is an Ontology Group Upregulated in Porcine Oocytes Before In Vitro Maturation: A Microarray Approach.

    Science.gov (United States)

    Budna, Joanna; Bryja, Artur; Celichowski, Piotr; Kranc, Wiesława; Ciesiółka, Sylwia; Borys, Sylwia; Rybska, Marta; Kolecka-Bednarczyk, Agata; Jeseta, Michal; Bukowska, Dorota; Antosik, Paweł; Brüssow, Klaus P; Bruska, Małgorzata; Nowicki, Michał; Zabel, Maciej; Kempisty, Bartosz

    2017-08-01

    Mammalian cumulus-oocyte complexes (COCs) reach full developmental capability during folliculogenesis and oogenesis. It is well recognized that only gametes achieving MII stage after in vivo or in vitro maturation (IVM) are successfully fertilized by a single spermatozoon. Although the process of oocyte nuclear and/or cytoplasmic maturation in pigs is well determined, there exist many differences that promote these processes in vivo and in vitro. Therefore, this study aimed to investigate the differences in RNA expression profiles between porcine oocytes before and after IVM using microarray and real-time quantitative polymerase chain reaction (RT-qPCR) assays. Experiments were performed on oocytes isolated from 55 pubertal crossbred Landrace gilts. The oocytes were analyzed both before and after IVM and only Brilliant Cresyl Blue (BCB)-positive gametes were used for subsequent microarray analysis (Affymetrix) and RT-qPCR analysis. The microarray assay, which measures expression of 12,258 transcripts, revealed 419 differentially expressed transcripts in porcine oocytes, from which 379 were downregulated and 40 were upregulated before IVM compared to those analyzed after IVM. After DAVID analysis, we found eight different transcripts, including IHH, BMP1, WWTR1, CHRDL1, KLF10, EIF2AK3, MMP14, and STC1. Their expression is related to the "bone development" ontology group and was further subjected to hierarchical clusterization. Using RT-qPCR analysis, we confirmed the results of the microarray assay, showing increased expression of the eight genes in oocytes before IVM compared to oocytes after maturation in vitro. It has been suggested that "bone development" belongs to one ontological group involving genes substantially upregulated in porcine oocytes before IVM. We suggest that the gamete mRNA expression profile before IVM may comprise stored transcripts, which are templates for protein biosynthesis following fertilization. We also hypothesize that these mRNAs may

  20. Technological Innovations for High-Throughput Approaches to In Vitro Allergy Diagnosis.

    Science.gov (United States)

    Chapman, Martin D; Wuenschmann, Sabina; King, Eva; Pomés, Anna

    2015-07-01

    Allergy diagnostics is being transformed by the advent of in vitro IgE testing using purified allergen molecules, combined with multiplex technology and biosensors, to deliver discriminating, sensitive, and high-throughput molecular diagnostics at the point of care. Essential elements of IgE molecular diagnostics are purified natural or recombinant allergens with defined purity and IgE reactivity, planar or bead-based multiplex systems to enable IgE to multiple allergens to be measured simultaneously, and, most recently, nanotechnology-based biosensors that facilitate rapid reaction rates and delivery of test results via mobile devices. Molecular diagnostics relies on measurement of IgE to purified allergens, the "active ingredients" of allergenic extracts. Typically, this involves measuring IgE to multiple allergens which is facilitated by multiplex technology and biosensors. The technology differentiates between clinically significant cross-reactive allergens (which could not be deduced by conventional IgE assays using allergenic extracts) and provides better diagnostic outcomes. Purified allergens are manufactured under good laboratory practice and validated using protein chemistry, mass spectrometry, and IgE antibody binding. Recently, multiple allergens (from dog) were expressed as a single molecule with high diagnostic efficacy. Challenges faced by molecular allergy diagnostic companies include generation of large panels of purified allergens with known diagnostic efficacy, access to flexible and robust array or sensor technology, and, importantly, access to well-defined serum panels form allergic patients for product development and validation. Innovations in IgE molecular diagnostics are rapidly being brought to market and will strengthen allergy testing at the point of care.

  1. Bayesian approach to model CD137 signaling in human M. tuberculosis in vitro responses.

    Directory of Open Access Journals (Sweden)

    Darío A Fernández Do Porto

    Full Text Available Immune responses are qualitatively and quantitatively influenced by a complex network of receptor-ligand interactions. Among them, the CD137:CD137L pathway is known to modulate innate and adaptive human responses against Mycobacterium tuberculosis. However, the underlying mechanisms of this regulation remain unclear. In this work, we developed a Bayesian Computational Model (BCM of in vitro CD137 signaling, devised to fit previously gathered experimental data. The BCM is fed with the data and the prior distribution of the model parameters and it returns their posterior distribution and the model evidence, which allows comparing alternative signaling mechanisms. The BCM uses a coupled system of non-linear differential equations to describe the dynamics of Antigen Presenting Cells, Natural Killer and T Cells together with the interpheron (IFN-γ and tumor necrosis factor (TNF-α levels in the media culture. Fast and complete mixing of the media is assumed. The prior distribution of the parameters that describe the dynamics of the immunological response was obtained from the literature and theoretical considerations Our BCM applies successively the Levenberg-Marquardt algorithm to find the maximum a posteriori likelihood (MAP; the Metropolis Markov Chain Monte Carlo method to approximate the posterior distribution of the parameters and Thermodynamic Integration to calculate the evidence of alternative hypothesis. Bayes factors provided decisive evidence favoring direct CD137 signaling on T cells. Moreover, the posterior distribution of the parameters that describe the CD137 signaling showed that the regulation of IFN-γ levels is based more on T cells survival than on direct induction. Furthermore, the mechanisms that account for the effect of CD137 signaling on TNF-α production were based on a decrease of TNF-α production by APC and, perhaps, on the increase in APC apoptosis. BCM proved to be a useful tool to gain insight on the mechanisms of CD

  2. Investigation of drug loading and in vitro release mechanisms of insulin-lauryl sulfate complex loaded PLGA nanoparticles.

    Science.gov (United States)

    Shi, K; Cui, F; Yamamoto, H; Kawashima, Y

    2008-12-01

    Insulin, a water soluble peptide hormone, was hydrophobically ion-paired with sodium lauryl sulfate (SDS) at the stoichiometric molar ratio of 6:1. The obtained insulin-SDS complex precipitation was subsequently formulated in biodegradable poly (D,L-lactic-co-glycolic acid) (PLGA) nanoparticles by a modified spontaneous emulsion solvent diffusion method. Compared with a conventional method for free insulin encapsulation, direct dissolution of SDS-paired insulin in the non-aqueous organic phase led to an increase in drug recovery from 42.5% to 89.6%. The more hydrophobic complex contributes to the improved affinity of insulin to the polymer matrix, resulting in a higher drug content in the nanoparticles. The drug loading was investigated by determining initial burst release at the first 30 min. The results showed that 64.8% of recovered drug were preferentially surface bound on complex loaded nanoparticles. The in vitro drug release was characterized by an initial burst and subsequent delayed release in dissolution media of deionized water and phosphate buffer saline (PBS). Compared with that in PBS, nanoparticles in deionized water medium presented very low initial burst release (15% vs. 65%) and incomplete cumulative release (25% vs. 90%) of the drug. In addition, dialysis experiments were performed to clarify the form of the released insulin in the dissolution media. The results suggested that the ion-pair complex was sensitive to ionic strength, insulin was released from the particular matrix as complex form and subsequently suffered dissociation from SDS in buffer saline. Moreover, the in vivo bioactivity of the SDS-paired insulin and nanoparticulate formulations were evaluated in mice by estimation of their blood sugar levels. The results showed that the bioactivity of insulin was unaltered after the ion-pairing process.

  3. In vitro model systems to investigate bile salt export pump (BSEP) activity and drug interactions: A review.

    Science.gov (United States)

    Cheng, Yaofeng; Woolf, Thomas F; Gan, Jinping; He, Kan

    2016-08-05

    The bile salt export pump protein (BSEP), expressed on the canalicular membranes of hepatocytes, is primarily responsible for the biliary excretion of bile salts. The inhibition of BSEP transport activity can lead to an increase in intracellular bile salt levels and liver injury. This review discusses the various in vitro assays currently available for assessing the effect of drugs or other chemical entities to modulate BSEP transport activity. BSEP transporter assays use one of the following platforms: Xenopus laevis oocytes; canalicular membrane vesicles (CMV); BSEP-expressed membrane vesicles; cell lines expressing BSEP; sandwich cultured hepatocytes (SCH); and hepatocytes in suspension. Two of these, BSEP-expressed insect membrane vesicles and sandwich cultured hepatocytes, are the most commonly used assays. BSEP membrane vesicles prepared from transfected insect cells are useful for assessing BSEP inhibition or substrate specificity and exploring mechanisms of BSEP-associated genetic diseases. This model can be applied in a high-throughput format for discovery-drug screening. However, experimental results from use of membrane vesicles may lack physiological relevance and the model does not allow for investigation of in situ metabolism in modulation of BSEP activity. Hepatocyte-based assays that use the SCH format provide results that are generally more physiologically relevant than membrane assays. The SCH model is useful in detailed studies of the biliary excretion of drugs and BSEP inhibition, but due to the complexity of SCH preparation, this model is used primarily for determining biliary clearance and BSEP inhibition in a limited number of compounds. The newly developed hepatocyte in suspension assay avoids many of the complexities of the SCH method. The use of pooled cryopreserved hepatocytes in suspension minimizes genetic variance and individual differences in BSEP activity and also provides the opportunity for higher throughput screening and cross

  4. The catechol-O-methyltransferase inhibitory potential of Z-vallesiachotamine by in silicoand in vitro approaches

    Directory of Open Access Journals (Sweden)

    Carolina dos Santos Passos

    Full Text Available AbstractZ-Vallesiachotamine is a monoterpene indole alkaloid that has a β-N-acrylate group in its structure. This class of compounds has already been described in different Psychotriaspecies. Our research group observed that E/Z-vallesiachotamine exhibits a multifunctional feature, being able to inhibit targets related to neurodegeneration, such as monoamine oxidase A, sirtuins 1 and 2, and butyrylcholinesterase enzymes. Aiming at better characterizing the multifunctional profile of this compound, its effect on cathecol-O-methyltransferase activity was investigated. The cathecol-O-methyltransferase activity was evaluated in vitro by a fluorescence-based method, using S-(5′-adenosyl-l-methionine as methyl donor and aesculetin as substrate. The assay optimization was performed varying the concentrations of methyl donor (S-(5′-adenosyl-l-methionine and enzyme. It was observed that the highest concentrations of both factors (2.25 U of the enzyme and 100 µM of S-(5′-adenosyl-l-methionine afforded the more reproducible results. The in vitro assay demonstrated that Z-vallesiachotamine was able to inhibit the cathecol-O-methyltransferase activity with an IC50 close to 200 µM. Molecular docking studies indicated that Z-vallesiachotamine can bind the catechol pocket of catechol-O-methyltransferase enzyme. The present work demonstrated for the first time the inhibitory properties of Z-vallesiachotamine on cathecol-O-methyltransferase enzyme, affording additional evidence regarding its multifunctional effects in targets related to neurodegenerative diseases.

  5. Anatomic investigation of lumbar transforaminal fenestration approach and its clinical application in far lateral disc herniation.

    Science.gov (United States)

    Xu, Baoshan; Xu, Haiwei; Destandau, Jean; Ma, Xinlong; He, Jian; Xia, Qun; Ji, Ning; Yang, Qiang; Liu, Yue; Jiang, Hongfeng

    2017-07-01

    Treating lumbar disease at the intervertebal foramina is controversial because we lack an approach providing sufficient exposure and preserving spinal stability. The primary objective of this study was to investigate the feasibility of the transforaminal fenestration (TFF) approach for treating lumbar disease involving the intervertebal foramina.In the anatomic study of 30 adult cadaveric lumbar spine specimens, the TFF approach was used from L1 to S1. The scope of resection was measured manually and on 3D CT images. 3D CT images of the lumbar spine of 31 adult patients were collected, and the scope of resection needed during the TFF approach was defined and measured from L1 to S1. In total, 30 patients (14 men) with lumbar FLDH underwent microendoscopic discectomy (MED) via the TFF approach. The results were evaluated with visual analog scale (VAS), Oswestry Disability Index (ODI), Short Form-36 survey (SF-36), and the MacNab scale.In cadavers, the TFF approach provided sufficient exposure to the posterolateral aspect of the disc and the exiting nerve root at all segments. At L1 to L4, a relatively small part of the isthmus and facet joint was resected; at L4 to S1, a relatively large part of the isthmus and facet joint was resected, so luniform fenestration was needed to preserve a more inferior articular process and continuity of the isthmus. Treatment with MED via the TFF approach was successful in the 30 patients with significantly relieved of symptom after operation. In total, 24 patients were followed for a mean of 24 months. The VAS, ODI, SF-36 physical component, and mental component summary scores at the final follow-up improved significantly compared with preoperative data (P approach can provide sufficient exposure to the lumbar intervertebal foramina with preservation of stability and can be used effectively with MED to treat lumbar FLDH.

  6. In Vitro Evaluation of New Approach to Digital Dental Model Articulation

    Science.gov (United States)

    Chang, Yu-Bing; Xia, James J.; Gateno, Jaime; Xiong, Zixiang; Teichgraeber, John F.; Lasky, Robert E.; Zhou, Xiaobo

    2015-01-01

    Purpose The purpose of the present study was to evaluate the accuracy of our newly developed approach to digital dental model articulation. Materials and Methods Twelve sets of stone dental models from patients with craniomaxillofacial deformities were used for validation. All the models had stable occlusion and no evidence of early contact. The stone models were hand articulated to the maximal intercuspation (MI) position and scanned using a 3-dimensional surface laser scanner. These digital dental models at the MI position served as the control group. To establish an experimental group, each mandibular dental model was disarticulated from its original MI position to 80 initial positions. Using a regular office personal computer, they were digitally articulated to the MI position using our newly developed approach. These rearticulated mandibular models served as the experimental group. Finally, the translational, rotational, and surface deviations in the mandibular position were calculated between the experimental and control groups, and statistical analyses were performed. Results All the digital dental models were successfully articulated. Between the control and experimental groups, the largest translational difference in mandibular position was within 0.2 mm ± 0.6 mm. The largest rotational difference was within 0.1° ± 1.1°. The averaged surface deviation was 0.08 ± 0.07. The results of the Bland and Altman method of assessing measurement agreement showed tight limits for the translational, rotational, and surface deviations. In addition, the final positions of the mandibular articulated from the 80 initial positions were absolutely agreed on. Conclusion The results of our study have demonstrated that using our approach, the digital dental models can be accurately and effectively articulated to the MI position. In addition, the 3-dimensional surface geometry of the mandibular teeth played a more important role in digital dental articulation than the

  7. On Goal-Oriented, Hydrogeological Site Investigation: A Holistic Approach (Henry Darcy Medal Lecture)

    Science.gov (United States)

    Rubin, Yoram

    2016-04-01

    UQ (for Uncertainty Quantification) is a critical element of groundwater management and by extension, of hydrological site investigation. While it is clear that UQ is an important goal, there is ambiguity as to what the target of the UQ should be, and how to make UQ relevant in the context of public policy. Planning for UQ (meaning what measurements to take, where, how many, what frequency, etc.), one could consider environmental performance parameters (EPMs, such as concentrations or travel time) as the targets of site investigation. But there is a need to go beyond EPMs, and to consider the uncertainty related to impacts such as enhanced cancer-risk due to groundwater contamination or, more generally, to decisions facing regulators. In any case, UQ requires site investigation, and decision-makers, who end up paying for it, are not really interested in EPMs: they care about making operational decisions that are defensible legally and justified from the perspective of public good. The key to UQ, whether considering EPMS or operational decisions concerning the public good, is defining a suitable strategy for site investigation. There is a body of published works on relating site investigations with EPMs, but much less is known on how to support operational decisions with strategies for site characterization. In this lecture, I will address this issue and I will outline a comprehensive approach for addressing it using a statistical formalism that couples hypothesis testing with Bayesian statistics. I refer to this approach as goal-oriented site investigation. I will show how site investigation strategies, with specifics such as which measurements to take and where, could be related to goals lined with operational decisions. This includes (1) defining the relevant goals; (2) formulating hypotheses; (3) defining alternative strategies for site investigation and (4) evaluating them in terms of probabilities for making errors in accepting or rejecting the hypotheses.

  8. Tumor Microenvironment In Experimental Models Of Human Cancer: Morphological Investigational Approaches

    Directory of Open Access Journals (Sweden)

    Lucia Minoli

    2017-05-01

    Discussion and conclusions. Due to the microenvironmental heterogeneity which influence tumor development and biological behavior, a sole quantification is unreliable for characterizing the TME. Considering that, morphological techniques proved to be a valuable approach, allowing the evaluation of the spatial distribution and mutual interaction between the different elements. Additional studies are needed for further investigate the biological significance of spatial distribution of the components of the TME.

  9. Comparative transcriptomic approach to investigate differences in wine yeast physiology and metabolism during fermentation.

    Science.gov (United States)

    Rossouw, Debra; Olivares-Hernandes, Roberto; Nielsen, Jens; Bauer, Florian F

    2009-10-01

    Commercial wine yeast strains of the species Saccharomyces cerevisiae have been selected to satisfy many different, and sometimes highly specific, oenological requirements. As a consequence, more than 200 different strains with significantly diverging phenotypic traits are produced globally. This genetic resource has been rather neglected by the scientific community because industrial strains are less easily manipulated than the limited number of laboratory strains that have been successfully employed to investigate fundamental aspects of cellular biology. However, laboratory strains are unsuitable for the study of many phenotypes that are of significant scientific and industrial interest. Here, we investigate whether a comparative transcriptomics and phenomics approach, based on the analysis of five phenotypically diverging industrial wine yeast strains, can provide insights into the molecular networks that are responsible for the expression of such phenotypes. For this purpose, some oenologically relevant phenotypes, including resistance to various stresses, cell wall properties, and metabolite production of these strains were evaluated and aligned with transcriptomic data collected during alcoholic fermentation. The data reveal significant differences in gene regulation between the five strains. While the genetic complexity underlying the various successive stress responses in a dynamic system such as wine fermentation reveals the limits of the approach, many of the relevant differences in gene expression can be linked to specific phenotypic differences between the strains. This is, in particular, the case for many aspects of metabolic regulation. The comparative approach therefore opens new possibilities to investigate complex phenotypic traits on a molecular level.

  10. Prediction of in-vivo pharmacokinetic profile for immediate and modified release oral dosage forms of furosemide using an in-vitro-in-silico-in-vivo approach.

    Science.gov (United States)

    Otsuka, Keiichi; Wagner, Christian; Selen, Arzu; Dressman, Jennifer

    2015-05-01

    To develop a physiologically based pharmacokinetic (PBPK) model for furosemide immediate release (IR) tablets and modified release (MR) capsules by coupling biorelevant dissolution testing results with pharmacokinetic (PK) and physiologic parameters, and to investigate the key factors influencing furosemide absorption using simulation approaches and the PBPK model. Using solubility, dissolution kinetics, gastrointestinal (GI) parameters and disposition parameters, a PBPK model for furosemide was developed with STELLA software. Solubility and dissolution profiles for both formulations were evaluated in biorelevant and compendial media. The simulated plasma profiles were compared with in-vivo profiles using point estimates of area under plasma concentration-time curve, maximal concentration after the dose and time to maximal concentration after the dose. Simulated plasma profiles of both furosemide IR tablets and MR capsules were similar to the observed in-vivo profile in terms of PK parameters. Sensitivity analysis of the IR tablet model indicated that both the gastric emptying and absorption rate have an influence on the plasma profile. For the MR capsules, the sensitivity analysis suggested that the release rate in the small intestine, gastric emptying and the absorption rate all have an influence on the plasma profile. A predictive model to describe both IR and MR dosage forms containing furosemide was attained. Because sensitivity analysis of the model is able to identify key factors influencing the plasma profile, this in-vitro-in-silico-in-vivo approach could be a useful tool for facilitating formulation development of drug products. © 2015 Royal Pharmaceutical Society.

  11. Functional in vitro tension measurements of fascial tissue ? a novel modified superfusion approach

    OpenAIRE

    Schleip, R.; Klingler, W.; Wearing, S; Naylor, I.; Zuegel, M.; Hoppe, K. (Kai)

    2016-01-01

    Introduction: While two laboratory techniques are commonly used to assess the tensile properties of muscle tissue, emerging evidence suggests that the fascial components of these tissues also serve an active role in force generation. Hence, we investigated whether these techniques are sensitive for assessment of fascial micromechanics. Methods: Force measurements on dissected fascial tissue were performed either using the classical immersion organ bath or using an improved superfusion approac...

  12. Biologically optimized helium ion plans: calculation approach and its in vitro validation

    Science.gov (United States)

    Mairani, A.; Dokic, I.; Magro, G.; Tessonnier, T.; Kamp, F.; Carlson, D. J.; Ciocca, M.; Cerutti, F.; Sala, P. R.; Ferrari, A.; Böhlen, T. T.; Jäkel, O.; Parodi, K.; Debus, J.; Abdollahi, A.; Haberer, T.

    2016-06-01

    Treatment planning studies on the biological effect of raster-scanned helium ion beams should be performed, together with their experimental verification, before their clinical application at the Heidelberg Ion Beam Therapy Center (HIT). For this purpose, we introduce a novel calculation approach based on integrating data-driven biological models in our Monte Carlo treatment planning (MCTP) tool. Dealing with a mixed radiation field, the biological effect of the primary 4He ion beams, of the secondary 3He and 4He (Z  =  2) fragments and of the produced protons, deuterons and tritons (Z  =  1) has to be taken into account. A spread-out Bragg peak (SOBP) in water, representative of a clinically-relevant scenario, has been biologically optimized with the MCTP and then delivered at HIT. Predictions of cell survival and RBE for a tumor cell line, characterized by {{(α /β )}\\text{ph}}=5.4 Gy, have been successfully compared against measured clonogenic survival data. The mean absolute survival variation ({μΔ \\text{S}} ) between model predictions and experimental data was 5.3%  ±  0.9%. A sensitivity study, i.e. quantifying the variation of the estimations for the studied plan as a function of the applied phenomenological modelling approach, has been performed. The feasibility of a simpler biological modelling based on dose-averaged LET (linear energy transfer) has been tested. Moreover, comparisons with biophysical models such as the local effect model (LEM) and the repair-misrepair-fixation (RMF) model were performed. {μΔ \\text{S}} values for the LEM and the RMF model were, respectively, 4.5%  ±  0.8% and 5.8%  ±  1.1%. The satisfactorily agreement found in this work for the studied SOBP, representative of clinically-relevant scenario, suggests that the introduced approach could be applied for an accurate estimation of the biological effect for helium ion radiotherapy.

  13. Biomechanics of Hybrid Anterior Cervical Fusion and Artificial Disc Replacement in 3-Level Constructs: An In Vitro Investigation

    Science.gov (United States)

    Liao, Zhenhua; Fogel, Guy R.; Pu, Ting; Gu, Hongsheng; Liu, Weiqiang

    2015-01-01

    Background The ideal surgical approach for cervical disk disease remains controversial, especially for multilevel cervical disease. The purpose of this study was to investigate the biomechanics of the cervical spine after 3-level hybrid surgery compared with 3-level anterior cervical discectomy and fusion (ACDF). Material/Methods Eighteen human cadaveric spines (C2-T1) were evaluated under displacement-input protocol. After intact testing, a simulated hybrid construct or fusion construct was created between C3 to C6 and tested in the following 3 conditions: 3-level disc plate disc (3DPD), 3-level plate disc plate (3PDP), and 3-level plate (3P). Results Compared to intact, almost 65~80% of motion was successfully restricted at C3-C6 fusion levels (p0.05). 3PDP construct resulted in significant decrease of ROM at C3-C6 levels less than 3P (pbiomechanical advantages compared to fusion in normalizing motion. PMID:26529430

  14. A novel approach to quantifying ovarian cell lipid content and lipid accumulation in vitro by confocal microscopy in lean women undergoing ovarian stimulation for in vitro fertilization (IVF).

    Science.gov (United States)

    Singh, Prapti; Amin, Marli; Keller, Erica; Simerman, Ariel; Aguilera, Paul; Briton-Jones, Christine; Hill, David L; Abbott, David H; Chazenbalk, Gregorio; Dumesic, Daniel A

    2013-06-01

    To quantify intracellular lipid levels in cumulus cells (CCs) and mural granulosa cells (MGCs) of lean women undergoing gonadotropin therapy for in vitro fertilization (IVF), based upon different cell preparation methods. CCs and MGCs from 16 lean women undergoing ovarian stimulation for IVF were studied. Cells were pooled by cell type, with each type of cell separated into two groups for determination of initial lipid content (Method 1) and subsequent lipid accumulation in vitro (Method 2). Cells for initial lipid content were immediately fixed at the time of the oocyte retrieval with 4% paraformaldehyde in suspension, while those for subsequent lipid accumulation in vitro were cultured for 4 h with 5% fetal calf serum and then fixed. Cells were treated with lipid fluorescent dye BODIPY® FL C16 and nuclear marker DAPI. Intracellular lipid was quantified by confocal microscopy, using ImageJ software analysis. There was no significant effect of cell type (P = 0.2) or cell type-cell preparation method interaction (P = 0.8) on cell area (Method 1: CC 99.7 ± 5.1, MGC 132.8 ± 5.8; Method 2: CC 221.9 ± 30.4, MGC 265.1 ± 48.5 μm(2)). The mean area of all cells combined was significantly less for cells prepared by Method 1 (116.2 ± 4.9 μm(2)) vs. Method 2 (243.5 ± 22.5 μm(2), P vitro was significantly higher in CC (154.0 ± 9.1) than MGC (104.6 ± 9.9 fluorescence/cell area, P vitro (P vitro over time.

  15. An integrated approach for detecting embryotoxicity and developmental toxicity of environmental contaminants using in vitro alternative methods.

    Science.gov (United States)

    Sogorb, Miguel A; Pamies, David; de Lapuente, Joaquín; Estevan, Carmen; Estévez, Jorge; Vilanova, Eugenio

    2014-10-15

    The main available alternatives for testing embryotoxicity are cellular tests with stem cells and in vitro-ex vivo tests with embryos. In cellular tests, the most developed alternative is the embryonic stem cell test, while the most developed tests involving embryos are the zebrafish and whole embryo culture test. They are technically more complex than cellular tests, but offer the advantage of determining the expectable phenotypic alteration caused by the exposure. Many efforts are currently being made, basically through proteomic and genomic approaches, in order to obtain improvements in predictivity of these tests. Development is a very complex process, and it is highly unlikely that a single alternative test can yield satisfactory performance with all types of chemicals. We propose a step-wise approach where model complexity, and consequently technical skills and economical costs, gradually increase if needed. The first level would be run short cellular assays to detect effects in early differentiation stages. The second level would involve longer cellular embryotoxicity tests to search embryotoxicants that have an effect on late differentiation stages. The third stage would consider tests with embryos because they allow the determination of hazards based on molecular and morphological alterations, and not only on differentiating cells. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  16. Apta-nanosensor preparation and in vitro assay for rapid Diazinon detection using a computational molecular approach.

    Science.gov (United States)

    Jokar, Mahmoud; Safaralizadeh, Mohammad Hassan; Hadizadeh, Farzin; Rahmani, Fatemeh; Kalani, Mohamad Reza

    2017-02-01

    Aptamers (ss-DNA or ss-RNA), also known as artificial antibodies, have been selected in vitro median to bind target molecules with high affinity and selectivity. Diazinon is one of the most widely used organophosphorus insecticides in developing and underdeveloped countries as insecticide and acaricide. Diazinon is readily absorbed from the gastrointestinal system and rapidly distributed throughout the body. Thus, the design of clinical and laboratory diagnostics using nanobiosensors is necessary. A computational approach allows us to screen or rank receptor structure and predict interaction outcomes with a deeper understanding, and it is much more cost effective than laboratory attempts. In this research, the best sequence (high affinity bind Diazinon-ssDNA) was ranked among 12 aptamers isolated from SELEX experimentation. Docking results, as the first virtual screening stage and static technique, selected frequent conformation of each aptamer. Then, the quantity and quality of aptamer-Diazinon interaction were simulated using molecular dynamics as a mobility technique. RMSD, RMSF, radius of gyration, and the number of hydrogen bonds formed between Diazinon-aptamer were monitored to assess the quantity and quality of interactions. G-quadruplex DNA aptamer (DF20) showed to be a reliable candidate for Diazinon biosensing. The apta-nanosensor designed using simulation results allowed with linearity detection in the range of .141-.65 nM and a LOD of 17.903 nM, and it was validated using a computational molecular approach.

  17. In vitro/in silico approach in the development of simvastatin-loaded self-microemulsifying drug delivery systems.

    Science.gov (United States)

    Ćetković, Zora; Cvijić, Sandra; Vasiljević, Dragana

    2017-12-19

    The aims of this study were to formulate simvastatin (SV)-loaded self-microemulsifying drug delivery systems (SMEDDS), and explore the potential of these drug delivery systems to improve SV solubility, and also to identify the optimal place in the gastrointestinal (GI) tract for the release of SV using coupled in vitro/in silico approach. In comparison to other published results, this study considered the extensive pre-systemic clearance of SV, which could significantly decrease its systemic and hepatic bioavailability if SV is delivered into the small intestine. SV-loaded SMEDDS were formulated using various proportions of oils (PEG 300 oleic glycerides, propylene glycol monocaprylate, propylene glycol monolaurate), surfactant (PEG 400 caprylic/capric glycerides) and cosurfactant (polysorbate 80) and subjected to characterization, and physiologically-based pharmacokinetic (PBPK) modeling. According to the in vitro results, the selected SMEDDS consisted of 10.0% PEG 300 oleic glycerides, 67.5% PEG 400 caprylic/capric glycerides, and 22.5% polysorbate 80. The use of acid-resistant capsules filled with SV-loaded SMEDDS was found helpful in protecting the drug against early degradation in proximal parts of the GI tract, however, in silico simulations indicated that pH-controlled drug release system that dissolve in the distal parts of the intestine might further improve SV bioavailability (up to 7.20%). The obtained results suggested that combined strategy for the improvement of SV bioavailability should comprise solubility enhancement and delayed drug release. The developed SV-specific PBPK model could potentially be used to assess the influence of formulation factors on drug absorption and disposition when developing SV oral dosage forms.

  18. Using a Dialogical Approach to Examine Peer Feedback During Chemistry Investigative Task Discussion

    Science.gov (United States)

    Gan Joo Seng, Mark; Hill, Mary

    2014-10-01

    Peer feedback is an inherent feature of classroom collaborative learning. Students invariably turn to their peers for feedback when carrying out an investigative task, and this feedback is usually implicit, unstructured and may positively or negatively influence students' learning when they work on a task. This study explored the characteristics of verbal peer feedback during a collaborative investigative chemistry task involving New Zealand Year 13 students. During the planning stage of the students' investigation, the discussions of five pairs of students were recorded and then transcribed. Analysis of transcribed verbal data focused on interactions that involved peer feedback along two dimensions, interactive/non-interactive and dialogic/authoritative (Mortimer and Scott, 2003). The findings indicated that although students adopted a predominantly interactive/authoritative communicative approach, with peer feedback as confirmation or evaluation, they are also capable of a more interactive/dialogic exchange, characterised by elaborative peer feedback. We discuss how this dialogic perspective on peer feedback provides an alternative approach to the analysis and study of student-student interactions during science investigations. The findings should be interpreted in light of the limitations in terms of sample size, grouping and specificity of the coding scheme. Implications for teacher practice are discussed in relation to facilitating peer feedback discourse in the science classroom.

  19. Preliminary Investigation of Self-Reported Emotional Responses to Approaching and Receding Footstep Sounds in a Virtual Reality Context

    DEFF Research Database (Denmark)

    Sikström, Erik; Nilsson, Niels Christian; Nordahl, Rolf

    2013-01-01

    The emotional impact of approaching and receding sounds sources studies has previously been studied in seated laboratory experiments in with and without accompanying visual stimulus. This paper investigates the emotional responses to approaching and receding footstep sounds in an interactive...

  20. submitter Biologically optimized helium ion plans: calculation approach and its in vitro validation

    CERN Document Server

    Mairani, A; Magro, G; Tessonnier, T; Kamp, F; Carlson, D J; Ciocca, M; Cerutti, F; Sala, P R; Ferrari, A; Böhlen, T T; Jäkel, O; Parodi, K; Debus, J; Abdollahi, A; Haberer, T

    2016-01-01

    Treatment planning studies on the biological effect of raster-scanned helium ion beams should be performed, together with their experimental verification, before their clinical application at the Heidelberg Ion Beam Therapy Center (HIT). For this purpose, we introduce a novel calculation approach based on integrating data-driven biological models in our Monte Carlo treatment planning (MCTP) tool. Dealing with a mixed radiation field, the biological effect of the primary $^4$He ion beams, of the secondary $^3$He and $^4$He (Z  =  2) fragments and of the produced protons, deuterons and tritons (Z  =  1) has to be taken into account. A spread-out Bragg peak (SOBP) in water, representative of a clinically-relevant scenario, has been biologically optimized with the MCTP and then delivered at HIT. Predictions of cell survival and RBE for a tumor cell line, characterized by ${{(\\alpha /\\beta )}_{\\text{ph}}}=5.4$ Gy, have been successfully compared against measured clonogenic survival data. The mean ...

  1. Identifying endogenous neural stem cells in the adult brain in vitro and in vivo: novel approaches.

    Science.gov (United States)

    Rueger, Maria Adele; Androutsellis-Theotokis, Andreas

    2013-01-01

    In the 1960s, Joseph Altman reported that the adult mammalian brain is capable of generating new neurons. Today it is understood that some of these neurons are derived from uncommitted cells in the subventricular zone lining the lateral ventricles, and the dentate gyrus of the hippocampus. The first area generates new neuroblasts which migrate to the olfactory bulb, whereas hippocampal neurogenesis seems to play roles in particular types of learning and memory. A part of these uncommitted (immature) cells is able to divide and their progeny can generate all three major cell types of the nervous system: neurons, astrocytes, and oligodendrocytes; these properties define such cells as neural stem cells. Although the roles of these cells are not yet clear, it is accepted that they affect functions including olfaction and learning/memory. Experiments with insults to the central nervous system also show that neural stem cells are quickly mobilized due to injury and in various disorders by proliferating, and migrating to injury sites. This suggests a role of endogenous neural stem cells in disease. New pools of stem cells are being discovered, suggesting an even more important role for these cells. To understand these cells and to coax them to contribute to tissue repair it would be very useful to be able to image them in the living organism. Here we discuss advances in imaging approaches as well as new concepts that emerge from stem cell biology with emphasis on the interface between imaging and stem cells.

  2. New in silico and conventional in vitro approaches to advance HIV drug discovery and design.

    Science.gov (United States)

    Maga, Giovanni; Veljkovic, Nevena; Crespan, Emmanuele; Spadari, Silvio; Prljic, Jelena; Perovic, Vladimir; Glisic, Sanja; Veljkovic, Veljko

    2013-01-01

    Recently, the new concept of the long-range intermolecular interactions in biological systems has been proposed. Combined use of molecular modeling techniques and the screening techniques based on the long-range interaction concept could significantly improve and accelerate discovery of new HIV drugs. However, any hit identified in silico needs to be characterized with respect to its biological target by enzymatic studies. Combined use of the in silico screening and the enzymatic studies allows an efficient selection of new anti-HIV drugs. The focus of this article is on the in silico screening of molecular libraries for candidate new HIV drugs, which is based on the molecular descriptors determining the long-range interaction between the drugs and their therapeutic target. This article also reviews the techniques for enzyme kinetic studies which are required for optimization of in silico selected candidate anti-HIV drugs. The novel approach of combining in silico screening techniques with enzymatic studies enables the accurate measurement of the quantitative descriptors of ligand-enzyme interactions. This novel method is a powerful tool for new anti-HIV drug discovery which can also reduce the drug development costs.

  3. Discovery of in vitro antitubercular agents through in silico ligand-based approaches.

    Science.gov (United States)

    De Vita, Daniela; Pandolfi, Fabiana; Cirilli, Roberto; Scipione, Luigi; Di Santo, Roberto; Friggeri, Laura; Mori, Mattia; Fiorucci, Diego; Maccari, Giorgio; Arul Christopher, Robert Selwyne; Zamperini, Claudio; Pau, Valentina; De Logu, Alessandro; Tortorella, Silvano; Botta, Maurizio

    2016-10-04

    The development of new anti-tubercular agents represents a constant challenge mostly due to the insurgency of resistance to the currently available drugs. In this study, a set of 60 molecules were selected by screening the Asinex and the ZINC collections and an in house library by means of in silico ligand-based approaches. Biological assays in Mycobacterium tuberculosis H37Ra ATCC 25177 strain highlighted (±)-1-(4-chlorophenyl)-2-(1H-imidazol-1-yl)ethyl-4-(3,4-dichlorophenyl)piperazine-1-carboxylate (5i) and 3-(4-chlorophenyl)-5-(2,4-dimethylpyrimidin-5-yl)-2-methylpyrazolo[1.5-a]pyrimidin-7(4H)-one (42) as the most potent compounds, having a Minimum Inhibitory Concentration (MIC) of 4 and 2 μg/mL respectively. These molecules represent a good starting point for further optimization of effective anti-TB agents. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  4. A three-dimensional approach to in vitro culture of immune-related cells

    DEFF Research Database (Denmark)

    Hartmann, Sofie Bruun

    in culture and to measure cell activity such as IGRA, as described above. The traditional way of culturing cells are done using polystyrene (PS) plastic ware in flask-, Petri dish- or micro titer plate format. However, these artificial two dimensional (2D) surfaces on which the cells grow, has shown...... on the differentiation of porcine monocytes. Monocytes are immune cells of high plasticity, and thus we speculated that they might be sensitive to culture conditions. Indeed, monocytes differentiated into monocyte-derived DC (moDCs) when cultured conventionally (2D PS) in the presence of GM-CSF and IL-4, but adopted...... a macrophage-like gene expression profile when cultured on PDMS. Further it was found that 3D culturing resulted in increased activation of the monocyte-derived cells. The work in this thesis covers several aspects within primary cell culture, but central to the work is the investigation of 3D cell culture...

  5. The ethological approach as a new way of investigating behavioural health in the Arctic.

    Science.gov (United States)

    Tafforin, Carole

    2011-04-01

    From an interdisciplinary perspective, the goal of our research in human ethology is to investigate the adaptive dynamics of small groups in isolation and confinement with a particular emphasis on unexplored environments, from circumpolar missions on earth to interplanetary missions in space. The ethological observations were designed to monitor the polar teams on the Subantarctic islands, the Antarctic continent and the Arctic Ocean. The working hypothesis viewed the periodic changes in the groups' organization as optimal behavioural strategies in extreme living and working conditions. The general methodological feature is a quantitative description of observable events based on the motor activity of individuals, interactions and communications among individuals and spatial mapping in collective areas. We observed group organization, group disorganization and group reorganization over extended time periods in the polar stations. Cultural grouping and gender-based individualities were observed as well. Focusing on the Tara Expedition in the Arctic, we observed variations in spatial indicators, including inter-individual positions, and in temporal indicators, like collective times, as behavioural strategies for preventing the monotony of social life. The ethological approach, using non-invasive techniques of observation, description and quantification of spontaneous human behaviour, offers an innovative and complementary tool for sociocultural approaches, enhancing the knowledge of contemporary circumpolar micro-societies. With the changing of environmental context, the Arctic natives would undertake changes in their group organization for maintaining their behavioural health. Such social adaptation could be investigated with this new approach in the field.

  6. Probe molecules (PrM) approach in adverse outcome pathway (AOP) based high throughput screening (HTS): in vivo discovery for developing in vitro target methods

    Science.gov (United States)

    Efficient and accurate adverse outcome pathway (AOP) based high-throughput screening (HTS) methods use a systems biology based approach to computationally model in vitro cellular and molecular data for rapid chemical prioritization; however, not all HTS assays are grounded by rel...

  7. A time scaling approach to develop an in vitro-in vivo correlation (IVIVC) model using a convolution-based technique.

    Science.gov (United States)

    Costello, Cian; Rossenu, Stefaan; Vermeulen, An; Cleton, Adriaan; Dunne, Adrian

    2011-10-01

    In vitro-in vivo correlation (IVIVC) models prove very useful during drug formulation development, the setting of dissolution specifications and bio-waiver applications following post approval changes. A convolution-based population approach for developing an IVIVC has recently been proposed as an alternative to traditional deconvolution based methods, which pose some statistical concerns. Our aim in this study was to use a time-scaling approach using a convolution-based technique to successfully develop an IVIVC model for a drug with quite different in vitro and in vivo time scales. The in vitro and the in vivo data were longitudinal in nature with considerable between subject variation in the in vivo data. The model was successfully developed and fitted to the data using the NONMEM package. Model utility was assessed by comparing model-predicted plasma concentration-time profiles with the observed in vivo profiles. This comparison met validation criteria for both internal and external predictability as set out by the regulatory authorities. This study demonstrates that a time-scaling approach may prove useful when attempting to develop an IVIVC for data with the aforementioned properties. It also demonstrates that the convolution-based population approach is quite versatile and that it is capable of producing an IVIVC model with a big difference between the in vitro and in vivo time scales.

  8. Complex spatial dynamics of oncolytic viruses in vitro: mathematical and experimental approaches.

    Directory of Open Access Journals (Sweden)

    Dominik Wodarz

    Full Text Available Oncolytic viruses replicate selectively in tumor cells and can serve as targeted treatment agents. While promising results have been observed in clinical trials, consistent success of therapy remains elusive. The dynamics of virus spread through tumor cell populations has been studied both experimentally and computationally. However, a basic understanding of the principles underlying virus spread in spatially structured target cell populations has yet to be obtained. This paper studies such dynamics, using a newly constructed recombinant adenovirus type-5 (Ad5 that expresses enhanced jellyfish green fluorescent protein (EGFP, AdEGFPuci, and grows on human 293 embryonic kidney epithelial cells, allowing us to track cell numbers and spatial patterns over time. The cells are arranged in a two-dimensional setting and allow virus spread to occur only to target cells within the local neighborhood. Despite the simplicity of the setup, complex dynamics are observed. Experiments gave rise to three spatial patterns that we call "hollow ring structure", "filled ring structure", and "disperse pattern". An agent-based, stochastic computational model is used to simulate and interpret the experiments. The model can reproduce the experimentally observed patterns, and identifies key parameters that determine which pattern of virus growth arises. The model is further used to study the long-term outcome of the dynamics for the different growth patterns, and to investigate conditions under which the virus population eliminates the target cells. We find that both the filled ring structure and disperse pattern of initial expansion are indicative of treatment failure, where target cells persist in the long run. The hollow ring structure is associated with either target cell extinction or low-level persistence, both of which can be viewed as treatment success. Interestingly, it is found that equilibrium properties of ordinary differential equations describing the

  9. The fitness landscape of HIV-1 gag: advanced modeling approaches and validation of model predictions by in vitro testing.

    Science.gov (United States)

    Mann, Jaclyn K; Barton, John P; Ferguson, Andrew L; Omarjee, Saleha; Walker, Bruce D; Chakraborty, Arup; Ndung'u, Thumbi

    2014-08-01

    Viral immune evasion by sequence variation is a major hindrance to HIV-1 vaccine design. To address this challenge, our group has developed a computational model, rooted in physics, that aims to predict the fitness landscape of HIV-1 proteins in order to design vaccine immunogens that lead to impaired viral fitness, thus blocking viable escape routes. Here, we advance the computational models to address previous limitations, and directly test model predictions against in vitro fitness measurements of HIV-1 strains containing multiple Gag mutations. We incorporated regularization into the model fitting procedure to address finite sampling. Further, we developed a model that accounts for the specific identity of mutant amino acids (Potts model), generalizing our previous approach (Ising model) that is unable to distinguish between different mutant amino acids. Gag mutation combinations (17 pairs, 1 triple and 25 single mutations within these) predicted to be either harmful to HIV-1 viability or fitness-neutral were introduced into HIV-1 NL4-3 by site-directed mutagenesis and replication capacities of these mutants were assayed in vitro. The predicted and measured fitness of the corresponding mutants for the original Ising model (r = -0.74, p = 3.6×10-6) are strongly correlated, and this was further strengthened in the regularized Ising model (r = -0.83, p = 3.7×10-12). Performance of the Potts model (r = -0.73, p = 9.7×10-9) was similar to that of the Ising model, indicating that the binary approximation is sufficient for capturing fitness effects of common mutants at sites of low amino acid diversity. However, we show that the Potts model is expected to improve predictive power for more variable proteins. Overall, our results support the ability of the computational models to robustly predict the relative fitness of mutant viral strains, and indicate the potential value of this approach for understanding viral immune evasion, and

  10. The fitness landscape of HIV-1 gag: advanced modeling approaches and validation of model predictions by in vitro testing.

    Directory of Open Access Journals (Sweden)

    Jaclyn K Mann

    2014-08-01

    Full Text Available Viral immune evasion by sequence variation is a major hindrance to HIV-1 vaccine design. To address this challenge, our group has developed a computational model, rooted in physics, that aims to predict the fitness landscape of HIV-1 proteins in order to design vaccine immunogens that lead to impaired viral fitness, thus blocking viable escape routes. Here, we advance the computational models to address previous limitations, and directly test model predictions against in vitro fitness measurements of HIV-1 strains containing multiple Gag mutations. We incorporated regularization into the model fitting procedure to address finite sampling. Further, we developed a model that accounts for the specific identity of mutant amino acids (Potts model, generalizing our previous approach (Ising model that is unable to distinguish between different mutant amino acids. Gag mutation combinations (17 pairs, 1 triple and 25 single mutations within these predicted to be either harmful to HIV-1 viability or fitness-neutral were introduced into HIV-1 NL4-3 by site-directed mutagenesis and replication capacities of these mutants were assayed in vitro. The predicted and measured fitness of the corresponding mutants for the original Ising model (r = -0.74, p = 3.6×10-6 are strongly correlated, and this was further strengthened in the regularized Ising model (r = -0.83, p = 3.7×10-12. Performance of the Potts model (r = -0.73, p = 9.7×10-9 was similar to that of the Ising model, indicating that the binary approximation is sufficient for capturing fitness effects of common mutants at sites of low amino acid diversity. However, we show that the Potts model is expected to improve predictive power for more variable proteins. Overall, our results support the ability of the computational models to robustly predict the relative fitness of mutant viral strains, and indicate the potential value of this approach for understanding viral immune evasion

  11. A global analysis approach for investigating structural resilience in urban drainage systems.

    Science.gov (United States)

    Mugume, Seith N; Gomez, Diego E; Fu, Guangtao; Farmani, Raziyeh; Butler, David

    2015-09-15

    Building resilience in urban drainage systems requires consideration of a wide range of threats that contribute to urban flooding. Existing hydraulic reliability based approaches have focused on quantifying functional failure caused by extreme rainfall or increase in dry weather flows that lead to hydraulic overloading of the system. Such approaches however, do not fully explore the full system failure scenario space due to exclusion of crucial threats such as equipment malfunction, pipe collapse and blockage that can also lead to urban flooding. In this research, a new analytical approach based on global resilience analysis is investigated and applied to systematically evaluate the performance of an urban drainage system when subjected to a wide range of structural failure scenarios resulting from random cumulative link failure. Link failure envelopes, which represent the resulting loss of system functionality (impacts) are determined by computing the upper and lower limits of the simulation results for total flood volume (failure magnitude) and average flood duration (failure duration) at each link failure level. A new resilience index that combines the failure magnitude and duration into a single metric is applied to quantify system residual functionality at each considered link failure level. With this approach, resilience has been tested and characterised for an existing urban drainage system in Kampala city, Uganda. In addition, the effectiveness of potential adaptation strategies in enhancing its resilience to cumulative link failure has been tested. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  12. Quantitative Prediction of Human Renal Clearance and Drug-Drug Interactions of Organic Anion Transporter Substrates Using In Vitro Transport Data: A Relative Activity Factor Approach.

    Science.gov (United States)

    Mathialagan, Sumathy; Piotrowski, Mary A; Tess, David A; Feng, Bo; Litchfield, John; Varma, Manthena V

    2017-04-01

    Organic anion transporters (OATs) are important in the renal secretion, and thus, the clearance, of many drugs; and their functional change can result in pharmacokinetic variability. In this study, we applied transport rates measured in vitro using OAT-transfected human embryonic kidney cells to predict human renal secretory and total renal clearance of 31 diverse drugs. Selective substrates to OAT1 (tenofovir), OAT2 (acyclovir and ganciclovir), and OAT3 (benzylpenicillin, oseltamivir acid) were used to obtain relative activity factors (RAFs) for these individual transporters by relating in vitro transport clearance (after physiologic scaling) to in vivo secretory clearance. Using the estimated RAFs (0.64, 7.3, and 4.1, respectively, for OAT1, OAT2, and OAT3, respectively) and the in vitro active clearances, renal secretory clearance and total renal clearance were predicted with average fold errors (AFEs) of 1.89 and 1.40, respectively. The results show that OAT3-mediated transport play a predominant role in renal secretion for 22 of the 31 drugs evaluated. This mechanistic static approach was further applied to quantitatively predict renal drug-drug interactions (AFE ∼1.6) of the substrate drugs with probenecid, a clinical probe OAT inhibitor. In conclusion, the proposed in vitro-in vivo extrapolation approach is the first comprehensive attempt toward mechanistic modeling of renal secretory clearance based on routinely employed in vitro cell models. Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.

  13. Evaluation of OASIS QSAR Models Using ToxCast™ in Vitro Estrogen and Androgen Receptor Binding Data and Application in an Integrated Endocrine Screening Approach.

    Science.gov (United States)

    Bhhatarai, Barun; Wilson, Daniel M; Price, Paul S; Marty, Sue; Parks, Amanda K; Carney, Edward

    2016-09-01

    Integrative testing strategies (ITSs) for potential endocrine activity can use tiered in silico and in vitro models. Each component of an ITS should be thoroughly assessed. We used the data from three in vitro ToxCast™ binding assays to assess OASIS, a quantitative structure-activity relationship (QSAR) platform covering both estrogen receptor (ER) and androgen receptor (AR) binding. For stronger binders (described here as AC50 75%) and specificity (> 86%) for ER as well as both high sensitivity (92-100%) and specificity (70-81%) for AR. For compounds within the domains of the ER and AR QSAR models that bound with AC50 approach wherein a) QSAR is used to identify compounds in-domain of the ER or AR binding models and predicted to bind; b) those compounds are screened in vitro to assess binding potency; and c) the stronger binders (AC50 vitro, that require metabolism to manifest activity, or for which in vivo AR testing is in order, need to be assessed differently. Bhhatarai B, Wilson DM, Price PS, Marty S, Parks AK, Carney E. 2016. Evaluation of OASIS QSAR models using ToxCast™ in vitro estrogen and androgen receptor binding data and application in an integrated endocrine screening approach. Environ Health Perspect 124:1453-1461; http://dx.doi.org/10.1289/EHP184.

  14. Large object investigation by digital holography with effective spectrum multiplexing under single-exposure approach

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ning, E-mail: coolboy006@sohu.com; Zhang, Yingying; Xie, Jun [College of Physics and Electronics, Nanjing XiaoZhuang University, Nanjing, Jiangsu Province 211171 (China)

    2014-10-13

    We present a method to investigate large object by digital holography with effective spectrum multiplexing under single-exposure approach. This method splits the original reference beam and redirects one of its branches as a second object beam. Through the modified Mach-Zehnder interferometer, the two object beams can illuminate different parts of the large object and create a spectrum multiplexed hologram onto the focal plane array of the charge-coupled device/complementary metal oxide semiconductor camera. After correct spectrum extraction and image reconstruction, the large object can be fully observed within only one single snap-shot. The flexibility and great performance make our method a very attractive and promising technique for large object investigation under common 632.8 nm illumination.

  15. A qualitative and quantitative investigation into the effect of fluoride formulations on enamel erosion and erosion–abrasion in vitro

    National Research Council Canada - National Science Library

    Austin, R.S; Stenhagen, K.S; Hove, L.H; Dunne, S; Moazzez, R; Bartlett, D.W; Tveit, A.B

    2011-01-01

    ... and protect enamel against dental erosion. In vitro topical application of a neutral 9500 ppm NaF solution induced CaF 2 like precipitates which resulted in penetration of enamel by sub-surface precipitates up to a depth of 40 μm. 3 Stannous fluoride (SnF 2 ) solutions can form a surface glaze or coating when applied to enamel, which could have result...

  16. An electrical impedance tomography-based approach to monitor in vitro sodium chloride dissolution from pharmaceutical tablets

    Science.gov (United States)

    Rimpiläinen, Ville; Heikkinen, Lasse M.; Kuosmanen, Marko; Lehikoinen, Anssi; Voutilainen, Arto; Vauhkonen, Marko; Ketolainen, Jarkko

    2009-10-01

    An approach to monitor in vitro dissolution process from pharmaceutical tablets utilizing electrical impedance tomography (EIT) is introduced. In the demonstration, a tablet containing sodium chloride (NaCl) was dissolution tested using tap water as a dissolution medium within an apparatus similar to the United States Pharmacopoeia dissolution apparatus II. During the process, the three-dimensional sodium chloride concentration distribution was monitored with EIT measurements as a function of time. For EIT measurements, an array of electrodes was attached on the boundary of the dissolution vessel, a set of alternating electric currents was injected through the electrodes, and the resulting voltages were measured. With these data and by applying mathematical algorithms, an approximation for the spatial/temporal concentration distribution inside the vessel was computed. It was found that the computed distributions were relatively homogeneous. A NaCl release curve was computed by integrating the concentration distribution over the vessel volume, and the final value of the curve matched well with the reference point based on the weight loss of the tablet. Finally, EIT monitoring is suggested to be used for research and product development purposes.

  17. A proposed in vitro model for investigating the mechanisms of 'joint cracking': a short report of preliminary techniques and observations.

    Science.gov (United States)

    Fryer, Jerome Cj; Quon, Jeffrey A; Vann, Richard D

    2017-03-01

    Joint "cracking" is common but not a clearly understood audible phenomenon. In this brief report we propose an in-vitro model to potentially assist in revealing a mechanism for, and therefore source of, this phenomenon. Using a suction cup under tension and de-nucleated fluid to simulate synovial fluid, an audible release with intra-articular cavity formation was elicited. This was followed by a refractory period during which no audible crack could be elicited until the observed cavity had slowly reabsorbed back into the joint fluid. Conversely, if regular fluid containing pre-existing nuclei was used, a cavity formation occurred but with neither an audible release nor subsequent refractory period. With this simple in-vitro model, we were able to reproduce the characteristic audible release, cavity formation and related refractory period typically observed in related experiments in human joints. This simple in-vitro model may be of use in helping to discern both the timing and precise nature of other yet to be discerned mechanisms related to joint cracking.

  18. Identifying barriers to and facilitators of tuberculosis contact investigation in Kampala, Uganda: a behavioral approach.

    Science.gov (United States)

    Ayakaka, Irene; Ackerman, Sara; Ggita, Joseph M; Kajubi, Phoebe; Dowdy, David; Haberer, Jessica E; Fair, Elizabeth; Hopewell, Philip; Handley, Margaret A; Cattamanchi, Adithya; Katamba, Achilles; Davis, J Lucian

    2017-03-09

    The World Health Organization recommends routine household tuberculosis contact investigation in high-burden countries but adoption has been limited. We sought to identify barriers to and facilitators of TB contact investigation during its introduction in Kampala, Uganda. We collected cross-sectional qualitative data through focus group discussions and interviews with stakeholders, addressing three core activities of contact investigation: arranging household screening visits through index TB patients, visiting households to screen contacts and refer them to clinics, and evaluating at-risk contacts coming to clinics. We analyzed the data using a validated theory of behavior change, the Capability, Opportunity, and Motivation determine Behavior (COM-B) model, and sought to identify targeted interventions using the related Behavior Change Wheel implementation framework. We led seven focus-group discussions with 61 health-care workers, two with 21 lay health workers (LHWs), and one with four household contacts of newly diagnosed TB patients. We, in addition, performed 32 interviews with household contacts from 14 households of newly diagnosed TB patients. Commonly noted barriers included stigma, limited knowledge about TB among contacts, insufficient time and space in clinics for counselling, mistrust of health-center staff among index patients and contacts, and high travel costs for LHWs and contacts. The most important facilitators identified were the personalized and enabling services provided by LHWs. We identified education, persuasion, enablement, modeling of health-positive behaviors, incentivization, and restructuring of the service environment as relevant intervention functions with potential to alleviate barriers to and enhance facilitators of TB contact investigation. The use of a behavioral theory and a validated implementation framework provided a comprehensive approach for systematically identifying barriers to and facilitators of TB contact

  19. Critical approaches in physical therapy research: investigating the symbolic value of walking.

    Science.gov (United States)

    Gibson, Barbara E; Teachman, Gail

    2012-08-01

    Research using a critical social science perspective is uncommon in physiotherapy (PT) despite its potential advantages for investigating questions other approaches cannot address. Critical approaches can be used to expose ideas and concepts that are dominant, given, or taken-for-granted in practice in order to reflect on how "things could be otherwise." The purpose of this paper is to use an example of research examining the symbolic value of walking to outline the key features of critical research and its application to PT. The study drew from Pierre Bourdieu's sociology of practice to illuminate how socially ingrained notions of normality and disability are reflected in rehabilitation practices and affect parents and children with cerebral palsy. Dominant social assumptions about the value of walking are shown to shape individual choices and contribute to parental feelings of angst and doubt, and negative self-identities for children. The example reveals how critical approaches to research can be used to reveal the socio-political dimension of rehabilitation practice and address important research questions that have been largely neglected.

  20. Investigating statistical differences in connectivity patterns properties at single subject level: a new resampling approach.

    Science.gov (United States)

    Toppi, J; Anzolin, A; Petti, M; Cincotti, F; Mattia, D; Salinari, S; Babiloni, F; Astolfi, L

    2014-01-01

    Methods based on the multivariate autoregressive (MVAR) approach are commonly used for effective connectivity estimation as they allow to include all available sources into a unique model. To ensure high levels of accuracy for high model dimensions, all the observations are used to provide a unique estimation of the model, and thus of the network and its properties. The unavailability of a distribution of connectivity values for a single experimental condition prevents to perform statistical comparisons between different conditions at a single subject level. This is a major limitation, especially when dealing with the heterogeneity of clinical conditions presented by patients. In the present paper we proposed a novel approach to the construction of a distribution of connectivity in a single subject case. The proposed approach is based on small perturbations of the networks properties and allows to assess significant changes in brain connectivity indexes derived from graph theory. Its feasibility and applicability were investigated by means of a simulation study and an application to real EEG data.

  1. New Approach for Investigating Reaction Dynamics and Rates with Ab Initio Calculations.

    Science.gov (United States)

    Fleming, Kelly L; Tiwary, Pratyush; Pfaendtner, Jim

    2016-01-21

    Herein, we demonstrate a convenient approach to systematically investigate chemical reaction dynamics using the metadynamics (MetaD) family of enhanced sampling methods. Using a symmetric SN2 reaction as a model system, we applied infrequent metadynamics, a theoretical framework based on acceleration factors, to quantitatively estimate the rate of reaction from biased and unbiased simulations. A systematic study of the algorithm and its application to chemical reactions was performed by sampling over 5000 independent reaction events. Additionally, we quantitatively reweighed exhaustive free-energy calculations to obtain the reaction potential-energy surface and showed that infrequent metadynamics works to effectively determine Arrhenius-like activation energies. Exact agreement with unbiased high-temperature kinetics is also shown. The feasibility of using the approach on actual ab initio molecular dynamics calculations is then presented by using Car-Parrinello MD+MetaD to sample the same reaction using only 10-20 calculations of the rare event. Owing to the ease of use and comparatively low-cost of computation, the approach has extensive potential applications for catalysis, combustion, pyrolysis, and enzymology.

  2. An Investigation of Primary School Teachers’ PCK towards Science Subjects Using an Inquiry-Based Approach

    Directory of Open Access Journals (Sweden)

    Menşure ALKIŞ KÜÇÜKAYDIN

    2016-09-01

    Full Text Available In this study, the pedagogical content knowledge (PCK of four experienced primary school teachers was investigated within the “Let’s Solve the Riddle of Our Body Unit”. The PCK investigation adopted a learning approach based on inquiry, content representation and pedagogical and professional-experience repertoires (PaP-eRs, and interview forms were used as data collection tools. During the course of the research, the findings obtained from observations made during a total of 18 course hours formed the basic data source of the study. According to the results of the study, in which descriptive and content analysis were used concurrently, primary school teachers lack subject matter knowledge, do not interrogate the pre-knowledge of students and some misconceptions exist regarding about blood moves and exercise with pulse. Additionally, some deficiencies were detected in the curriculum, i.e., it offers non-inquisitional knowledge. Furthermore, teachers employee assessment methods with traditional teaching methods and techniques. In the context of an inquiry-based learning approach, teachers appeared to believe that classroom activities were adversely affected by the physical conditions (class size, lack of laboratory etc., students’ cognitive levels and parent profiles. The result of this study revealed that PCK components affect one another. The PCK findings pertaining to primary school teachers as it concerns the unit are briefly discussed and some suggestions about the development of PCK are submitted.

  3. Development of a Combined In Vitro Physiologically Based Kinetic (PBK) and Monte Carlo Modelling Approach to Predict Interindividual Human Variation in Phenol-Induced Developmental Toxicity.

    Science.gov (United States)

    Strikwold, Marije; Spenkelink, Bert; Woutersen, Ruud A; Rietjens, Ivonne M C M; Punt, Ans

    2017-06-01

    With our recently developed in vitro physiologically based kinetic (PBK) modelling approach, we could extrapolate in vitro toxicity data to human toxicity values applying PBK-based reverse dosimetry. Ideally information on kinetic differences among human individuals within a population should be considered. In the present study, we demonstrated a modelling approach that integrated in vitro toxicity data, PBK modelling and Monte Carlo simulations to obtain insight in interindividual human kinetic variation and derive chemical specific adjustment factors (CSAFs) for phenol-induced developmental toxicity. The present study revealed that UGT1A6 is the primary enzyme responsible for the glucuronidation of phenol in humans followed by UGT1A9. Monte Carlo simulations were performed taking into account interindividual variation in glucuronidation by these specific UGTs and in the oral absorption coefficient. Linking Monte Carlo simulations with PBK modelling, population variability in the maximum plasma concentration of phenol for the human population could be predicted. This approach provided a CSAF for interindividual variation of 2.0 which covers the 99th percentile of the population, which is lower than the default safety factor of 3.16 for interindividual human kinetic differences. Dividing the dose-response curve data obtained with in vitro PBK-based reverse dosimetry, with the CSAF provided a dose-response curve that reflects the consequences of the interindividual variability in phenol kinetics for the developmental toxicity of phenol. The strength of the presented approach is that it provides insight in the effect of interindividual variation in kinetics for phenol-induced developmental toxicity, based on only in vitro and in silico testing. © The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  4. Band gaps from the Tran-Blaha modified Becke-Johnson approach: a systematic investigation.

    Science.gov (United States)

    Jiang, Hong

    2013-04-07

    The semi-local Becke-Johnson (BJ) exchange-correlation potential and its modified form proposed by Tran and Blaha (TB-mBJ) have attracted a lot of interest recently because of the surprisingly accurate band gaps they can deliver for many semiconductors and insulators. In this work, we have investigated the performance of the TB-mBJ potential for the description of electronic band structures in a comprehensive set of semiconductors and insulators. We point out that a perturbative use of the TB-mBJ potential can give overall better results. By investigating a set of IIB-VI and III-V semiconductors, we point out that although the TB-mBJ approach can describe the band gap of these materials quite well, the binding energies of semi-core d-states in these materials deviate strongly from experiment. The difficulty of the TB-mBJ potential to describe the localized states is likely the cause for the fact that the electronic band structures of Cu2O and La2O3 are still poorly described. Based on these observations, we propose to combine the TB-mBJ approach with the Hubbard U correction for localized d∕f states, which is able to provide overall good descriptions for both the band gaps and semi-core states binding energies. We further apply the approach to calculate the band gaps of a set of Ti(IV)-oxides, many of which have complicated structures so that the more advanced methods like GW are expensive to treat directly. An overall good agreement with experiment is obtained, which is remarkable considering its little computational efforts compared to GW.

  5. Combining in vitro, in vivo and in silico approaches to evaluate nutraceutical potentials and chemical fingerprints of Moltkia aurea and Moltkia coerulea.

    Science.gov (United States)

    Zengin, Gokhan; Ceylan, Ramazan; Katanić, Jelena; Mollica, Adriano; Aktumsek, Abdurrahman; Boroja, Tatjana; Matić, Sanja; Mihailović, Vladimir; Stanić, Snežana; Aumeeruddy-Elalfi, Zaahira; Yilmaz, Mustafa Abdullah; Mahomoodally, Mohamad Fawzi

    2017-09-01

    Methanolic extracts of Moltkia aurea Boiss. (MA) and Moltkia coerulea (Willd.) Lehm. (MC) were investigated for their antioxidant capacity and enzymatic inhibitory potential against acetylcholinesterase, butyrylcholinesterase, α-amylase, α-glucosidase, and tyrosinase in vitro. MA and MC were also explored for their antimicrobial effect, as well as for their possible genotoxic/antigenotoxic potential on Drosophila melanogaster in vivo. The total bioactive components (phenolic (TPC) and flavonoid contents (TFC)) were determined and liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolite profiling of MA and MC appraised. The plausible docking poses of bioactive compounds to key enzymes were further studied using molecular modelling approach. MA proved to be a better antioxidant with higher TPC and TFC compared to MC. Protocatechuic acid, rutin, hesperidin and malic acid were the most abundant in these extracts. Both MA and MC exhibited antigenotoxic potential with a %R in DNA damage of 60.90 and 53.14% respectively. The docking studies revealed that rutin, hesperidin, and rosmarinic acid have the best scores for all the enzymes tested. MA and MC were found to be rich in phytochemicals with potent antioxidant, antimicrobial, and antigenotoxic activities that can be further studied for the management of neurodegenerative complications, diabetes, and hyperpigmentation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. In vitro and in vivo investigation on genomic stability of Salmonella enterica Typhimurium DT41 obtained from broiler breeders in Denmark

    DEFF Research Database (Denmark)

    Barua, Himel; Lindblom, Ina Lucilia; Bisgaard, Magne

    2013-01-01

    Salmonella enterica serovar Typhimurium phage type DT41 has previously been identified from salmonella-positive broiler breeder flocks in Denmark and isolates obtained from different flocks have demonstrated major diversity by multiple-locus variable-number tandem-repeats analysis (MLVA) typing...... and human origin, respectively. The in vitro experiment demonstrated that DT41 survived more than 6 months in feed-pellets at 20 °C whereas the survival in dust was less than 4 weeks. Infection pattern and excretion varied for the poultry and human strain and birds of different age groups as revealed...... by the in vivo experiment. Genetic stability of cultures obtained from the in vitro and in vivo survival/passage was investigated by plasmid profiling, pulsed-field gel electrophoresis (PFGE) and MLVA. The results of plasmid profiling and PFGE demonstrated genomic stability of all but one strain kept in dust...

  7. In vitro solubility, dissolution and permeability studies combined with semi-mechanistic modeling to investigate the intestinal absorption of desvenlafaxine from an immediate- and extended release formulation

    DEFF Research Database (Denmark)

    Franek, F; Jarlfors, A; Larsen, F.

    2015-01-01

    (ERF). Semi-mechanistic models of desvenlafaxine were built (using SimCyp®) by combining in vitro data on dissolution and permeation (mechanistic part of model) with clinical data (obtained from literature) on distribution and clearance (non-mechanistic part of model). The model predictions...... for desvenlafaxine absorption (i.e. intestinal dissolution or permeation) is not fully clarified. The aim of this study was to investigate whether dissolution and/or intestinal permeability rate-limit desvenlafaxine absorption from an immediate-release formulation (IRF) and Pristiq®, an extended release formulation...... of desvenlafaxine pharmacokinetics after IRF and ERF administration were compared with published clinical data from 14 trials. Desvenlafaxine in vivo dissolution from the IRF and ERF was predicted from in vitro solubility studies and biorelevant dissolution studies (using the USP3 dissolution apparatus...

  8. Enhancing the regeneration of bone defects by alkalizing the peri-implant zone – an in vitro approach

    Directory of Open Access Journals (Sweden)

    Galow Anne-Marie

    2016-09-01

    Full Text Available The effects of alkaline pH on the initial adhesion of osteoblasts to titanium surfaces was analyzed by single cell force microscopy (SCFM. In the SCFM measurements, the same cells were used to compare their unspecific adhesion to uncoated titanium with their specific adhesion to collagen coated titanium. When the maximum detachment forces (MDFs were compared at pH 7.4 and 8.0, only slight differences were found on pure titanium, while the MDFs were significantly increased at collagen coated surfaces at pH 8.0. Effects on the subsequent proliferation and gene expression were investigated in an in vitro model system consisting of an alkalizing polyvinyl alcohol (PVA matrix and a perforated titanium disc. The sodium hydroxide releasing matrix maintained the medium pH between pH 7.6 and pH 8.4 during the entire experiment. Under these conditions, cell counts were significantly increased with respect to the control system after 7 days in culture. These results were supported by gene expression analyses, which showed an upregulation of proliferation-controlling genes of the EGFR1 and PI3K/AKT pathways after 14 days in culture. The SCFM data were complemented by findings of an intensive regulation of genes known to be associated with focal adhesion such as Itga8 and Tnn.

  9. Studying inhibition of calcium oxalate stone formation: an in vitro approach for screening hydrogen sulfide and its metabolites

    Directory of Open Access Journals (Sweden)

    S. Vaitheeswari

    2015-06-01

    Full Text Available ABSTRACTPurpose:Calcium oxalate urolithiasis is one of the most common urinary tract diseases and is of high prevalence. The present study proposes to evaluate the antilithiatic property of hydrogen sulfide and its metabolites like thiosulfate & sulfate in an in vitro model.Materials and Methods:The antilithiatic activity of sodium hydrogen sulfide (NaSH, sodium thiosulfate (Na2S2O3 and sodium sulfate (Na2SO4 on the kinetics of calcium oxalate crystal formation was investigated both in physiological buffer and in urine from normal and recurrent stone forming volunteers. The stones were characterized by optical and spectroscopic techniques.Results:The stones were characterized to be monoclinic, prismatic and bipyramidal habit which is of calcium monohydrate and dihydrate nature. The FTIR displayed fingerprint corresponding to calcium oxalate in the control while in NaSH treated, S=O vibrations were visible in the spectrum. The order of percentage inhibition was NaSH>Na2S2O3>Na2SO4.Conclusion:Our study indicates that sodium hydrogen sulfide and its metabolite thiosulfate are inhibitors of calcium oxalate stone agglomeration which makes them unstable both in physiological buffer and in urine. This effect is attributed to pH changes and complexing of calcium by S2O32-and SO42- moiety produced by the test compounds.

  10. A combined three-dimensional in vitro-in silico approach to modelling bubble dynamics in decompression sickness.

    Science.gov (United States)

    Walsh, C; Stride, E; Cheema, U; Ovenden, N

    2017-12-01

    The growth of bubbles within the body is widely believed to be the cause of decompression sickness (DCS). Dive computer algorithms that aim to prevent DCS by mathematically modelling bubble dynamics and tissue gas kinetics are challenging to validate. This is due to lack of understanding regarding the mechanism(s) leading from bubble formation to DCS. In this work, a biomimetic in vitro tissue phantom and a three-dimensional computational model, comprising a hyperelastic strain-energy density function to model tissue elasticity, were combined to investigate key areas of bubble dynamics. A sensitivity analysis indicated that the diffusion coefficient was the most influential material parameter. Comparison of computational and experimental data revealed the bubble surface's diffusion coefficient to be 30 times smaller than that in the bulk tissue and dependent on the bubble's surface area. The initial size, size distribution and proximity of bubbles within the tissue phantom were also shown to influence their subsequent dynamics highlighting the importance of modelling bubble nucleation and bubble-bubble interactions in order to develop more accurate dive algorithms. © 2017 The Authors.

  11. Modulatory effect of three antibiotics on uterus bovine contractility in vitro and likely therapeutic approaches in reproduction.

    Science.gov (United States)

    Piccinno, M; Rizzo, A; Maselli, M A; Derosa, M; Sciorsci, R L

    2014-12-01

    This in vitro study investigates the modulatory effect of three antibiotics (amoxicillin, enrofloxacin, and rifaximin) on contractility of the bovine uterine tissue in follicular and luteal phases. The effects of these antibiotics at three single doses (10(-6), 10(-5), and 10(-4) M) on their basal contractility were evaluated in isolated organ bath. The functionality of the strip throughout the experiment was evaluated by a dose of carbachol (10(-5) M); the obtained effect had to be repeatable (difference of ≤20%) that is comparable to that induced by the previous administration of the same substance. The results demonstrate the different modulatory activities of these antibiotics on uterine contractility in follicular and luteal phases. The effects induced by amoxicillin and enrofloxacin are opposite: the first relaxes and the second increases the uterine contractility in both cycle phases. Instead, the activity of rifaximin varies depending on the phase of estrous cycle: it increases in the follicular phase and relaxes in the luteal phase. The obtained data provide the hypothesis of possible implications of these drugs in the pharmacologic modulation of uterine contractions. Their action at this level, associated with their specific antimicrobial effects, could suggest using these antibiotics for the treatment of diseases related to postpartum or infections that may occur in pregnant cattle, by virtue of their effects on myometrial contractility too. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Hybrid in Silico/in Vitro Approach for the Identification of Angiotensin I Converting Enzyme Inhibitory Peptides from Parma Dry-Cured Ham.

    Science.gov (United States)

    Dellafiora, Luca; Paolella, Sara; Dall'Asta, Chiara; Dossena, Arnaldo; Cozzini, Pietro; Galaverna, Gianni

    2015-07-22

    The bioactivity assessment of foodborne peptides is currently a research area of great relevance, and, in particular, several studies are devoted to the antihypertensive effects through the inhibition of angiotensin I converting enzyme (ACE). In the present work, a straightforward workflow to identify inhibitory peptides from food matrices is proposed, which involves a hybrid in vitro/in silico tandem approach. Parma dry-cured ham was chosen as case study. In particular, the advantage of using the hybrid approach to identify active sequences (in comparison to the experimental trials alone) has been pointed out. Specifically, fractions obtained by in vitro gastrointestinal digestion of ham samples of 18 and 24 months of aging have been assessed for ACE inhibition. At the same time, the released peptidomic profiles, which cannot be entirely evaluated by using in vitro assays, have been screened for the inhibition by using an in silico model. Then, to identify novel inhibitory sequences, a series of strong candidates have been synthesized and assessed for their inhibitory activity through in vitro assay. On the one hand, the use of computational simulations appeared to be an effective strategy to find active sequences, as confirmed by in vitro analysis. On the other hand, strong inhibitory sequences were identified for the first time in Parma dry-cured ham (e.g., LGL and SFVTT with IC50 values of 145 and 395 μM, respectively), which is a product of international dietary and economic relevance. Therefore, these findings demonstrate the usefulness of in silico methodologies coupled to in vitro tests for the identification of potentially bioactive peptides, and they give an important contribution to the study of the overall nutritional value of Parma ham.

  13. Methodological factors affecting gas and methane production during in vitro rumen fermentation evaluated by meta-analysis approach.

    Science.gov (United States)

    Maccarana, Laura; Cattani, Mirko; Tagliapietra, Franco; Schiavon, Stefano; Bailoni, Lucia; Mantovani, Roberto

    2016-01-01

    Effects of some methodological factors on in vitro measures of gas production (GP, mL/g DM), CH4 production (mL/g DM) and proportion (% CH4 on total GP) were investigated by meta-analysis. These factors were considered: pressure in the GP equipment (0 = constant; 1 = increasing), incubation time (0 = 24; 1 = ≥ 48 h), time of rumen fluid collection (0 = before feeding; 1 = after feeding of donor animals), donor species of rumen fluid (0 = sheep; 1 = bovine), presence of N in the buffer solution (0 = presence; 1 = absence), and ratio between amount of buffered rumen fluid and feed sample (BRF/FS; 0 = ≤ 130 mL/g DM; 1 = 130-140 mL/g DM; 2 = ≥ 140 mL/g DM). The NDF content of feed sample incubated (NDF) was considered as a continuous variable. From an initial database of 105 papers, 58 were discarded because one of the above-mentioned factors was not stated. After discarding 17 papers, the final dataset comprised 30 papers (339 observations). A preliminary mixed model analysis was carried out on experimental data considering the study as random factor. Variables adjusted for study effect were analyzed using a backward stepwise analysis including the above-mentioned variables. The analysis showed that the extension of incubation time and reduction of NDF increased GP and CH4 values. Values of GP and CH4 also increased when rumen fluid was collected after feeding compared to before feeding (+26.4 and +9.0 mL/g DM, for GP and CH4), from bovine compared to sheep (+32.8 and +5.2 mL/g DM, for GP and CH4), and when the buffer solution did not contain N (+24.7 and +6.7 mL/g DM for GP and CH4). The increase of BRF/FS ratio enhanced GP and CH4 production (+7.7 and +3.3 mL/g DM per each class of increase, respectively). In vitro techniques for measuring GP and CH4 production are mostly used as screening methods, thus a full standardization of such techniques is not feasible. However, a greater harmonization

  14. Application of Model-Based Approaches to Evaluate Hepatic Transporter-Mediated Drug Clearance: In vitro, In vivo, and In vitro-In vivo Extrapolation.

    Science.gov (United States)

    Liu, Zhihao; Liu, Kexin

    2016-01-01

    Hepatic transporters, including efflux transporters and uptake transporters, have been recognized to play an important role in the disposition of various drugs. These membrane transporters show extensive substrate specificity with an abundance of overlap, implying the probability of transporter involved in the drug-drug interactions with other drugs. Thus reliable techniques are taken into consideration to evaluate the role of transporter-mediated drug disposition and possible effects on pharmacokinetics in the research. An electronic search of PubMed database from inception to December, 2015 was conducted. In addition, we searched the reference lists of included studies and carried out a citation search for the included studies via Web of Science to find other potentially relevant studies. The function of membrane transporters could be evaluated in vitro, in situ and in vivo, employing models spanning from cell-based assay to transgenic mouse. Each technique has its own application with specific advantages and limitations. The readers will gain insight into techniques applied to evaluate the drug clearance mediated by hepatic transporters. In addition, this review focuses particularly on the in vitro-in vivo extrapolation of hepatic transporter-mediated drug clearance. The challenges and gaps of the extrapolation are further discussed. The increased understanding of this knowledge would improve the capability to predict the in vivo situation.

  15. Human in vitro 3D co-culture model to engineer vascularized bone-mimicking tissues combining computational tools and statistical experimental approach.

    Science.gov (United States)

    Bersini, Simone; Gilardi, Mara; Arrigoni, Chiara; Talò, Giuseppe; Zamai, Moreno; Zagra, Luigi; Caiolfa, Valeria; Moretti, Matteo

    2016-01-01

    The generation of functional, vascularized tissues is a key challenge for both tissue engineering applications and the development of advanced in vitro models analyzing interactions among circulating cells, endothelium and organ-specific microenvironments. Since vascularization is a complex process guided by multiple synergic factors, it is critical to analyze the specific role that different experimental parameters play in the generation of physiological tissues. Our goals were to design a novel meso-scale model bridging the gap between microfluidic and macro-scale studies, and high-throughput screen the effects of multiple variables on the vascularization of bone-mimicking tissues. We investigated the influence of endothelial cell (EC) density (3-5 Mcells/ml), cell ratio among ECs, mesenchymal stem cells (MSCs) and osteo-differentiated MSCs (1:1:0, 10:1:0, 10:1:1), culture medium (endothelial, endothelial + angiopoietin-1, 1:1 endothelial/osteo), hydrogel type (100%fibrin, 60%fibrin+40%collagen), tissue geometry (2 × 2 × 2, 2 × 2 × 5 mm(3)). We optimized the geometry and oxygen gradient inside hydrogels through computational simulations and we analyzed microvascular network features including total network length/area and vascular branch number/length. Particularly, we employed the "Design of Experiment" statistical approach to identify key differences among experimental conditions. We combined the generation of 3D functional tissue units with the fine control over the local microenvironment (e.g. oxygen gradients), and developed an effective strategy to enable the high-throughput screening of multiple experimental parameters. Our approach allowed to identify synergic correlations among critical parameters driving microvascular network development within a bone-mimicking environment and could be translated to any vascularized tissue. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Once-daily propranolol extended-release tablet dosage form: formulation design and in vitro/in vivo investigation.

    Science.gov (United States)

    Huang, Yaw-Bin; Tsai, Yi-Hung; Yang, Wan-Chiech; Chang, Jui-Sheng; Wu, Pao-Chu; Takayama, Kozo

    2004-11-01

    The purpose of this study was to develop and optimize the propranolol once-daily extended release formulations containing HPMC, Microcrystalline cellulose (MCC) and lactose. In vitro studies, the response surface methodology and multiple response optimization utilizing the polynomial equation were used to search for the optimal formulation with specific release rate at different time intervals. The constrained mixture experimental design was used to prepare systematic model formulations, which were composed of three formulation variables: the content of HPMC (X(1)) MCC (X(2)) and lactose (X(3)). The drug release percent at 1.5, 4, 8, 14 and 24 h were the target responses and were restricted to 15-30, 35-55, 55-75, 75-90 and 90-110%, respectively. The results showed that the optimized formulation provided a dissolution pattern equivalent to the predicted curve, which indicated that the optimal formulation could be obtained using response surface methodology. The mechanism of drug release from HMPC matrix tablets followed non-Fickian diffusion. In the vivo study, the MRT was prolonged for matrix tablets when compared with commercial immediate release tablets. Furthermore, a linear relationship between in vitro dissolution and in vivo absorption was observed in the beagle dogs.

  17. Investigation of approaches for hydrogeophysical joint inversion using a parallel computing platform

    Science.gov (United States)

    Commer, M.; Kowalsky, M. B.; Doetsch, J.; Newman, G. A.; Finsterle, S.

    2012-12-01

    Owing to the computational demands of joint inverse problems, we have developed a parallel inversion framework based on the inverse modeling tool iTOUGH2. The software provides inverse modeling capabilities for TOUGH2, a general-purpose simulator for multiphase, multicomponent, non-isothermal flows in multidimensional fractured-porous media. Our parallel version of iTOUGH2 realizes a hierarchically parallel architecture using the Message Passing Interface. This architecture allows for large numbers of parallel processes to be employed for addressing many-parameter inverse problems on large meshes. To provide geophysical modeling capabilities, we have further combined iTOUGH2 with the geophysical simulator Electromagnetic Geological Mapper (EMGeo), which features a suite of methods for modeling electrical and electromagnetic data types, including controlled-source electromagnetics, magnetotellurics, electrical resistivity tomography, and (spectral) induced polarization. Here, we consider the estimation of hydrological parameters through inverse modeling of hydrological and geophysical data associated with experiments from the shallow unconfined uranium-contaminated aquifer at the DOE Integrated Field Research Challenge (IFRC) site at Rifle, Colorado. Our study draws on examples from ongoing research at the IFRC site, and investigates different approaches for jointly inverting electrical resistivity tomography (ERT) and tracer concentration data. Exploiting complementary hydrological-geophysical modeling and parallel computing capabilities of the inversion framework, we investigate a few sequential and coupled inversion approaches, initially using synthetic examples. In the first approach, done as a pre-processing step before hydrological inversion, geophysical inversion of crosswell ERT data yields spatial maps of the subsurface electrical resistivity. During subsequent hydrological inversion, these maps are matched with their counterparts calculated from a

  18. Investigating students' view on STEM in learning about electrical current through STS approach

    Science.gov (United States)

    Tupsai, Jiraporn; Yuenyong, Chokchai

    2018-01-01

    This study aims to investigate Grade 11 students' views on Science Technology Engineering Mathematics (STEM) with the integration of learning about electrical current based on Science Technology Society (STS) approach [8]. The participants were 60 Grade 11 students in Demonstration Secondary School, Khon Kaen University, Khon Kaen Province, Thailand. The methodology is in the respect of interpretive paradigm. The teaching and learning about Electrical Current through STS approach carried out over 6 weeks. The Electrical Current unit through STS approach was developed based on framework[8] that consists of five stages including (1) identification of social issues, (2) identification of potential solutions, (3) need for knowledge, (4) decision making, and (5) socialization stage. To start with, the question "what if this world is lack of electricity" was challenged in the class in order to move students to find the problem of how to design Electricity Generation from Clean Energy. Students were expected to apply scientific and other knowledge to design of Electricity Generation. Students' views on STEM were collected during their learning by participant' observation and students' tasks. Their views on STEM were categorized when they applied their knowledge for designing the Electricity Generation. The findings indicated that students cooperatively work to solve the problem when applying knowledge about the content of Science and Mathematics and processing skill of Technology and Engineering. It showed that students held the integration of science, technology, engineering and mathematics to design their possible solutions in learning about Electrical Current. The paper also discusses implications for science teaching and learning through STS in Thailand.

  19. [Applicability of the da Vinci robotic system in the skull base surgical approach. Preclinical investigation].

    Science.gov (United States)

    Fernandez-Nogueras Jimenez, Francisco J; Segura Fernandez-Nogueras, Miguel; Jouma Katati, Majed; Arraez Sanchez, Miguel Ángel; Roda Murillo, Olga; Sánchez Montesinos, Indalecio

    2015-01-01

    The role of robotic surgery is well established in various specialties such as urology and general surgery, but not in others such as neurosurgery and otolaryngology. In the case of surgery of the skull base, it has just emerged from an experimental phase. To investigate possible applications of the da Vinci surgical robot in transoral skull base surgery, comparing it with the authors' experience using conventional endoscopic transnasal surgery in the same region. A transoral transpalatal approach to the nasopharynx and medial skull base was performed on 4 cryopreserved cadaver heads. We used the da Vinci robot, a 30° standard endoscope 12mm thick, dual camera and dual illumination, Maryland forceps on the left terminal and curved scissors on the right, both 8mm thick. Bone drilling was performed manually. For the anatomical study of this region, we used 0.5cm axial slices from a plastinated cadaver head. Various skull base structures at different depths were reached with relative ease with the robot terminals Transoral robotic surgery with the da Vinci system provides potential advantages over conventional endoscopic transnasal surgery in the surgical approach to this region. Copyright © 2014 Sociedad Española de Neurocirugía. Published by Elsevier España. All rights reserved.

  20. Investigating articulated heavy-vehicle crashes in Western Australia using a spatial approach.

    Science.gov (United States)

    Gudes, Ori; Varhol, Richard; Sun, Qian Chayn; Meuleners, Lynn

    2017-09-01

    Recent developments in Western Australia's economy including widespread traffic congestion as well as road safety issues are increasingly becoming prominent. Previous studies relied on traditional statistical methods to investigate patterns and characteristics of motor vehicle crashes. Although useful, statistical analysis alone is incapable of providing a spatial context and is therefore unable to associate existing crash characteristics with a spatial distribution. To identify concentrations or "hotspots" of articulated heavy vehicle crashes in WA between the years 2001-2013, by using a spatial analysis approach. Spatial modelling and spatio-temporal analytical methods such as Emerging Hotspots were used to identify emerging hotspots on specific roads in Western Australia using the Integrated Road Information System (IRIS). The results suggest that the majority of articulated heavy vehicles crashes occurred in the vicinity or within the Perth metropolitan area. Based on spatial-temporal trend analyses, our findings highlight some regions that are emerging as areas of interest. This study was one of the first attempts to adopt a spatial analysis approach in studying heavy-vehicle crashes in Western Australia. Applying spatial methodologies to road safety data has the potential of obtaining previously undiscovered insights, which can be extended further, and provide future avenues to research in this field. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Investigation of approach slab and its settlement for roads and bridges.

    Science.gov (United States)

    2014-01-01

    Approach slabs serve as a transitional system between an approach road and a bridge. Settlement of bridge approach slabs and their : supporting backfill has been experienced by more than ten Departments of Transportation throughout the United States....

  2. Functional and molecular responses of the blue mussel Mytilus edulis' hemocytes exposed to cadmium - An in vitro model and transcriptomic approach.

    Science.gov (United States)

    Granger Joly de Boissel, Philippine; Fournier, Michel; Rodriguez-Lecompte, Juan Carlos; McKenna, Patty; Kibenge, Frederick; Siah, Ahmed

    2017-08-01

    The bivalve mollusk, Mytilus edulis, is used as a sentinel species in several monitoring programs due to its ability to bio-accumulate contaminants. Its immune system consists of hemocytes and humoral components, which constitute the main part of the hemolymph. The present study is aimed at understanding the effects of Cd on the differentially expressed genes involved in the phagocytosis of M. edulis' hemocytes. Our approach focuses on an in vitro model by exposing hemocytes to different concentrations of Cd ranging from 10-9 M to 10-3 M. Phagocytosis and cell viability as functional markers were measured using flow cytometry. The molecular mechanisms regulated by Cd were investigated using RNA-seq and DGE analysis. Results showed that viability and phagocytosis of hemocytes exposed to 10-3 M of Cd were significantly decreased after 21 h of exposure. RNA sequencing data showed that 1112 transcripts (out of 352,976 contigs) were differentially regulated by the highest concentration of Cd. Among these identified transcripts, 1028 and 84 were up and down-regulated respectively. The induction of super oxide dismutase (SOD), glutathion-s-transferase (GST), cytochrome P450 2C8 (CYP2C8), multidrug resistance protein (MRP1) and heat shock protein 70 (HSP70) suggests that Cd can regulate key molecular mechanisms. In addition, several toll-like receptors (TLR) as well as genes involved in phagocytosis (actin and CDC42) and apoptosis (caspase 8 and XIAP/IAP) were induced by Cd. Thus, our model highlights the effect of Cd on the phagocytic function of M. edulis' hemocytes along with the regulation of gene expression involved in innate immunity, detoxification and apoptosis. Further investigations need to be pursued to unravel the effects of Cd on the molecular mechanisms identified in this study. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. An integrated approach for prospectively investigating a mode-of-action for rodent liver effects

    Energy Technology Data Exchange (ETDEWEB)

    LeBaron, Matthew J., E-mail: MJLeBaron@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Geter, David R., E-mail: dave.geter@gmail.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Rasoulpour, Reza J. [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Gollapudi, B. Bhaskar, E-mail: BBGollapudi@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Thomas, Johnson, E-mail: JThomas4@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Murray, Jennifer, E-mail: AMurray@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Kan, H. Lynn, E-mail: HLKan@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Wood, Amanda J., E-mail: AJWood@dow.com [Toxicology and Environmental Research and Consulting, The Dow Chemical Company, Midland, MI, 48674 (United States); Elcombe, Cliff, E-mail: CliffElcombe@cxrbiosciences.com [CXR Biosciences, 2 James Lindsay Place, Dundee Technopole, Dundee, DD1 5JJ, Scotland (United Kingdom); Vardy, Audrey, E-mail: audrey_vardy@europe.bd.com [CXR Biosciences, 2 James Lindsay Place, Dundee Technopole, Dundee, DD1 5JJ, Scotland (United Kingdom); McEwan, Jillian, E-mail: jillian.mcewan@rtmcewan.co.uk [CXR Biosciences, 2 James Lindsay Place, Dundee Technopole, Dundee, DD1 5JJ, Scotland (United Kingdom); Terry, Claire, E-mail: CTerry@dow.com [Dow AgroSciences, Abingdon, Oxfordshire (United Kingdom); Billington, Richard, E-mail: RBillington@dow.com [Dow AgroSciences, Abingdon, Oxfordshire (United Kingdom)

    2013-07-15

    Registration of new plant protection products (e.g., herbicide, insecticide, or fungicide) requires comprehensive mammalian toxicity evaluation including carcinogenicity studies in two species. The outcome of the carcinogenicity testing has a significant bearing on the overall human health risk assessment of the substance and, consequently, approved uses for different crops across geographies. In order to understand the relevance of a specific tumor finding to human health, a systematic, transparent, and hypothesis-driven mode of action (MoA) investigation is, appropriately, an expectation by the regulatory agencies. Here, we describe a novel approach of prospectively generating the MoA data by implementing additional end points to the standard guideline toxicity studies with sulfoxaflor, a molecule in development. This proactive MoA approach results in a more robust integration of molecular with apical end points while minimizing animal use. Sulfoxaflor, a molecule targeting sap-feeding insects, induced liver effects (increased liver weight due to hepatocellular hypertrophy) in an initial palatability probe study for selecting doses for subsequent repeat-dose dietary studies. This finding triggered the inclusion of dose-response investigations of the potential key events for rodent liver carcinogenesis, concurrent with the hazard assessment studies. As predicted, sulfoxaflor induced liver tumors in rats and mice in the bioassays. The MoA data available by the time of the carcinogenicity finding supported the conclusion that the carcinogenic potential of sulfoxaflor was due to CAR/PXR nuclear receptor activation with subsequent hepatocellular proliferation. This MoA was not considered to be relevant to humans as sulfoxaflor is unlikely to induce hepatocellular proliferation in humans and therefore would not be a human liver carcinogen. - Highlights: • We prospectively generated MoA data into standard guideline toxicity studies. • A proactive MoA approach

  4. In vitro Staphylococcus aureus–induced oxidative stress in mice murine peritoneal macrophages: a duration–dependent approach

    Directory of Open Access Journals (Sweden)

    Subhankari Prasad Chakraborty

    2014-05-01

    Conclusions: From this study, it may be summarized that in vitro VSSA infection not only generates excess free radical but also affects the antioxidant status and glutathione cycle in murine peritoneal macrophages.

  5. mRNA Expression is a Relevant Tool to Identify Developmental Neurotoxicants Using an In Vitro Approach

    National Research Council Canada - National Science Library

    Hogberg, Helena T; Kinsner-Ovaskainen, Agnieszka; Coecke, Sandra; Hartung, Thomas; Bal-Price, Anna K

    ...) is based entirely on in vivo studies that are both time consuming and costly, there is a need to develop alternative in vitro methods for initial screening to prioritize chemicals for further DNT testing...

  6. Biopolymer-mediated synthesis of Fe3O4 nanoparticles and investigation of their in vitro cytotoxicity effects

    Science.gov (United States)

    Gholoobi, Aida; Meshkat, Zahra; Abnous, Khalil; Ghayour-Mobarhan, Majid; Ramezani, Mohammad; Homaei Shandiz, Fatemeh; Verma, K. D.; Darroudi, Majid

    2017-08-01

    Superparamagnetic iron oxide nanoparticles (SPIONs; Fe3O4) were synthesized by a ;green; co-precipitation method in aqueous starch solution as a food media. Powder X-ray diffraction (PXRD) patterns indicated that the synthesized samples were pure Fe3O4 with a spinel structure, and the coating of starch did not undergo any phase change. Fourier transform infrared (FTIR) spectra confirmed the formation of starch coated Fe3O4 nanoparticles. Field emission scanning electron microscopy (FESEM) micrographs illustrated the formation of nanoparticles in the size range of below 25 nm. Magnetic measurements revealed that the saturated magnetization of the starch-SPIONs reached 36.5 emu/g. The non-toxic effect of SPIONs concentration below 50 and 100 μg/ml was observed in the studies of in vitro cytotoxicity on normal and cancerous cell lines, respectively. The dose dependent toxicity made it a suitable candidate for various medical applications.

  7. A systematic in vitro investigation on poly-arginine modified nanostructured lipid carrier: Pharmaceutical characteristics, cellular uptake, mechanisms and cytotoxicity

    Directory of Open Access Journals (Sweden)

    Mingshuang Sun

    2017-01-01

    Full Text Available The aim of the present study was to develop a poly-arginine modified nanostructured lipid carrier (R-NLC by fusion-emulsification method and to test its pharmaceutical characteristics. The influence of R-NLC on A549 cells like cellular uptake and cytotoxicity was also appraised using unmodified NLC as the controlled group. As the results revealed, R-NLC had an average diameter of about 40 nm and a positive zeta potential of about +17 mv, the entrapment efficiency decreased apparently, and no significant difference on the in vitro drug release was found after R8-modification. The cellular uptake and cytotoxicity increased obviously compared with unmodified NLC. The cellular uptake mechanisms of R-NLC involved energy, macropinocytosis, clathrin-mediated endocytosis, and caveolin-mediated endocytosis. The outcomes of the present study strongly support the theory that cell penetrating peptides have the ability of enhancing the cellular uptake of nanocarriers.

  8. Investigation of sonar transponders for offshore wind farms: modeling approach, experimental setup, and results.

    Science.gov (United States)

    Fricke, Moritz B; Rolfes, Raimund

    2013-11-01

    The installation of offshore wind farms in the German Exclusive Economic Zone requires the deployment of sonar transponders to prevent collisions with submarines. The general requirements for these systems have been previously worked out by the Research Department for Underwater Acoustics and Marine Geophysics of the Bundeswehr. In this article, the major results of the research project "Investigation of Sonar Transponders for Offshore Wind Farms" are presented. For theoretical investigations a hybrid approach was implemented using the boundary element method to calculate the source directivity and a three-dimensional ray-tracing algorithm to estimate the transmission loss. The angle-dependence of the sound field as well as the weather-dependence of the transmission loss are compared to experimental results gathered at the offshore wind farm alpha ventus, located 45 km north of the island Borkum. While theoretical and experimental results are in general agreement, the implemented model slightly underestimates scattering at the rough sea surface. It is found that the source level of 200 dB re 1 μPa at 1 m is adequate to satisfy the detectability of the warning sequence at distances up to 2 NM (≈3.7 km) within a horizontal sector of ±60° if realistic assumptions about signal-processing and noise are made. An arrangement to enlarge the angular coverage is discussed.

  9. Investigation of the scaling characteristics of LANDSAT temperature and vegetation data: a wavelet-based approach.

    Science.gov (United States)

    Rathinasamy, Maheswaran; Bindhu, V M; Adamowski, Jan; Narasimhan, Balaji; Khosa, Rakesh

    2017-10-01

    An investigation of the scaling characteristics of vegetation and temperature data derived from LANDSAT data was undertaken for a heterogeneous area in Tamil Nadu, India. A wavelet-based multiresolution technique decomposed the data into large-scale mean vegetation and temperature fields and fluctuations in horizontal, diagonal, and vertical directions at hierarchical spatial resolutions. In this approach, the wavelet coefficients were used to investigate whether the normalized difference vegetation index (NDVI) and land surface temperature (LST) fields exhibited self-similar scaling behaviour. In this study, l-moments were used instead of conventional simple moments to understand scaling behaviour. Using the first six moments of the wavelet coefficients through five levels of dyadic decomposition, the NDVI data were shown to be statistically self-similar, with a slope of approximately -0.45 in each of the horizontal, vertical, and diagonal directions of the image, over scales ranging from 30 to 960 m. The temperature data were also shown to exhibit self-similarity with slopes ranging from -0.25 in the diagonal direction to -0.20 in the vertical direction over the same scales. These findings can help develop appropriate up- and down-scaling schemes of remotely sensed NDVI and LST data for various hydrologic and environmental modelling applications. A sensitivity analysis was also undertaken to understand the effect of mother wavelets on the scaling characteristics of LST and NDVI images.

  10. Investigation of the scaling characteristics of LANDSAT temperature and vegetation data: a wavelet-based approach

    Science.gov (United States)

    Rathinasamy, Maheswaran; Bindhu, V. M.; Adamowski, Jan; Narasimhan, Balaji; Khosa, Rakesh

    2017-10-01

    An investigation of the scaling characteristics of vegetation and temperature data derived from LANDSAT data was undertaken for a heterogeneous area in Tamil Nadu, India. A wavelet-based multiresolution technique decomposed the data into large-scale mean vegetation and temperature fields and fluctuations in horizontal, diagonal, and vertical directions at hierarchical spatial resolutions. In this approach, the wavelet coefficients were used to investigate whether the normalized difference vegetation index (NDVI) and land surface temperature (LST) fields exhibited self-similar scaling behaviour. In this study, l-moments were used instead of conventional simple moments to understand scaling behaviour. Using the first six moments of the wavelet coefficients through five levels of dyadic decomposition, the NDVI data were shown to be statistically self-similar, with a slope of approximately -0.45 in each of the horizontal, vertical, and diagonal directions of the image, over scales ranging from 30 to 960 m. The temperature data were also shown to exhibit self-similarity with slopes ranging from -0.25 in the diagonal direction to -0.20 in the vertical direction over the same scales. These findings can help develop appropriate up- and down-scaling schemes of remotely sensed NDVI and LST data for various hydrologic and environmental modelling applications. A sensitivity analysis was also undertaken to understand the effect of mother wavelets on the scaling characteristics of LST and NDVI images.

  11. Evaluation of two dynamic in vitro models simulating fasted and fed state conditions in the upper gastrointestinal tract (TIM-1 and tiny-TIM) for investigating the bioaccessibility of pharmaceutical compounds from oral dosage forms

    NARCIS (Netherlands)

    Verwei, M.; Minekus, M.; Zeijdner, E.; Schilderink, R.; Havenaar, R.

    2016-01-01

    Pharmaceutical research needs predictive in vitro tools for API bioavailability in humans. We evaluated two dynamic in vitro gastrointestinal models: TIM-1 and tiny-TIM. Four low-soluble APIs in various formulations were investigated in the TIM systems under fasted and fed conditions. API

  12. Magnetic Particle / Magnetic Resonance Imaging: In-Vitro MPI-Guided Real Time Catheter Tracking and 4D Angioplasty Using a Road Map and Blood Pool Tracer Approach.

    Science.gov (United States)

    Salamon, Johannes; Hofmann, Martin; Jung, Caroline; Kaul, Michael Gerhard; Werner, Franziska; Them, Kolja; Reimer, Rudolph; Nielsen, Peter; Vom Scheidt, Annika; Adam, Gerhard; Knopp, Tobias; Ittrich, Harald

    2016-01-01

    In-vitro evaluation of the feasibility of 4D real time tracking of endovascular devices and stenosis treatment with a magnetic particle imaging (MPI) / magnetic resonance imaging (MRI) road map approach and an MPI-guided approach using a blood pool tracer. A guide wire and angioplasty-catheter were labeled with a thin layer of magnetic lacquer. For real time MPI a custom made software framework was developed. A stenotic vessel phantom filled with saline or superparamagnetic iron oxide nanoparticles (MM4) was equipped with bimodal fiducial markers for co-registration in preclinical 7T MRI and MPI. In-vitro angioplasty was performed inflating the balloon with saline or MM4. MPI data were acquired using a field of view of 37.3×37.3×18.6 mm3 and a frame rate of 46 volumes/sec. Analysis of the magnetic lacquer-marks on the devices were performed with electron microscopy, atomic absorption spectrometry and micro-computed tomography. Magnetic marks allowed for MPI/MRI guidance of interventional devices. Bimodal fiducial markers enable MPI/MRI image fusion for MRI based roadmapping. MRI roadmapping and the blood pool tracer approach facilitate MPI real time monitoring of in-vitro angioplasty. Successful angioplasty was verified with MPI and MRI. Magnetic marks consist of micrometer sized ferromagnetic plates mainly composed of iron and iron oxide. 4D real time MP imaging, tracking and guiding of endovascular instruments and in-vitro angioplasty is feasible. In addition to an approach that requires a blood pool tracer, MRI based roadmapping might emerge as a promising tool for radiation free 4D MPI-guided interventions.

  13. Magnetic Particle / Magnetic Resonance Imaging: In-Vitro MPI-Guided Real Time Catheter Tracking and 4D Angioplasty Using a Road Map and Blood Pool Tracer Approach.

    Directory of Open Access Journals (Sweden)

    Johannes Salamon

    Full Text Available In-vitro evaluation of the feasibility of 4D real time tracking of endovascular devices and stenosis treatment with a magnetic particle imaging (MPI / magnetic resonance imaging (MRI road map approach and an MPI-guided approach using a blood pool tracer.A guide wire and angioplasty-catheter were labeled with a thin layer of magnetic lacquer. For real time MPI a custom made software framework was developed. A stenotic vessel phantom filled with saline or superparamagnetic iron oxide nanoparticles (MM4 was equipped with bimodal fiducial markers for co-registration in preclinical 7T MRI and MPI. In-vitro angioplasty was performed inflating the balloon with saline or MM4. MPI data were acquired using a field of view of 37.3×37.3×18.6 mm3 and a frame rate of 46 volumes/sec. Analysis of the magnetic lacquer-marks on the devices were performed with electron microscopy, atomic absorption spectrometry and micro-computed tomography.Magnetic marks allowed for MPI/MRI guidance of interventional devices. Bimodal fiducial markers enable MPI/MRI image fusion for MRI based roadmapping. MRI roadmapping and the blood pool tracer approach facilitate MPI real time monitoring of in-vitro angioplasty. Successful angioplasty was verified with MPI and MRI. Magnetic marks consist of micrometer sized ferromagnetic plates mainly composed of iron and iron oxide.4D real time MP imaging, tracking and guiding of endovascular instruments and in-vitro angioplasty is feasible. In addition to an approach that requires a blood pool tracer, MRI based roadmapping might emerge as a promising tool for radiation free 4D MPI-guided interventions.

  14. Interactions among biotic and abiotic factors affect the reliability of tungsten microneedles puncturing in vitro and in vivo peripheral nerves: A hybrid computational approach

    Energy Technology Data Exchange (ETDEWEB)

    Sergi, Pier Nicola, E-mail: p.sergi@sssup.it [Translational Neural Engineering Laboratory, The Biorobotics Institute, Scuola Superiore Sant' Anna, Viale Rinaldo Piaggio 34, Pontedera, 56025 (Italy); Jensen, Winnie [Department of Health Science and Technology, Fredrik Bajers Vej 7, 9220 Aalborg (Denmark); Yoshida, Ken [Department of Biomedical Engineering, Indiana University - Purdue University Indianapolis, 723 W. Michigan St., SL220, Indianapolis, IN 46202 (United States)

    2016-02-01

    Tungsten is an elective material to produce slender and stiff microneedles able to enter soft tissues and minimize puncture wounds. In particular, tungsten microneedles are used to puncture peripheral nerves and insert neural interfaces, bridging the gap between the nervous system and robotic devices (e.g., hand prostheses). Unfortunately, microneedles fail during the puncture process and this failure is not dependent on stiffness or fracture toughness of the constituent material. In addition, the microneedles' performances decrease during in vivo trials with respect to the in vitro ones. This further effect is independent on internal biotic effects, while it seems to be related to external biotic causes. Since the exact synergy of phenomena decreasing the in vivo reliability is still not known, this work explored the connection between in vitro and in vivo behavior of tungsten microneedles through the study of interactions between biotic and abiotic factors. A hybrid computational approach, simultaneously using theoretical relationships and in silico models of nerves, was implemented to model the change of reliability varying the microneedle diameter, and to predict in vivo performances by using in vitro reliability and local differences between in vivo and in vitro mechanical response of nerves. - Highlights: • We provide phenomenological Finite Element (FE) models of peripheral nerves to study the interactions with W microneedles • We provide a general interaction-based approach to model the reliability of slender microneedles • We evaluate the reliability of W microneedels to puncture in vivo nerves • We provide a novel synergistic hybrid approach (theory + simulations) involving interactions among biotic and abiotic factors • We validate the hybrid approach by using experimental data from literature.

  15. Probe molecule (PrM) approach in adverse outcome pathway (AOP) based high-throughput screening (HTS): in vivo discovery for developing in vitro target methods.

    Science.gov (United States)

    Angrish, Michelle M; Madden, Michael C; Pleil, Joachim D

    2015-04-20

    Efficient and accurate adverse outcome pathway (AOP) based high-throughput screening (HTS) methods use a systems biology based approach to computationally model in vitro cellular and molecular data for rapid chemical prioritization; however, not all HTS assays are grounded by relevant in vivo exposure data. The challenge is to develop HTS assays with unambiguous quantitative links between in vitro responses and corresponding in vivo effects, which is complicated by metabolically insufficient systems, in vitro to in vivo extrapolation (IVIVE), cross-species comparisons, and other inherent issues correlating IVIVE findings. This article introduces the concept of ultrasensitive gas phase probe molecules (PrMs) to help bridge the current HTS assay IVIVE gap. The PrM concept assesses metabolic pathways that have already been well-defined from intact human or mammalian models. Specifically, the idea is to introduce a gas phase probe molecule into a system, observe normal steady state, add chemicals of interest, and quantitatively measure (from headspace gas) effects on PrM metabolism that can be directly linked back to a well-defined and corresponding in vivo effect. As an example, we developed the pharmacokinetic (PK) parameters and differential equations to estimate methyl tertiary butyl ether (MTBE) metabolism to tertiary butyl alcohol (TBA) via cytochrome (CYP) 2A6 in the liver from human empirical data. Because MTBE metabolic pathways are well characterized from in vivo data, we can use it as a PrM to explore direct and indirect chemical effects on CYP pathways. The PrM concept could be easily applied to in vitro and alternative models of disease and phenotype, and even test for volatile chemicals while avoiding liquid handling robotics. Furthermore, a PrM can be designed for any chemical with known empirical human exposure data and used to assess chemicals for which no information exists. Herein, we propose an elegant gas phase probe molecule-based approach to in

  16. Pharmaceutical characterization of novel tenofovir liposomal formulations for enhanced oral drug delivery: in vitro pharmaceutics and Caco-2 permeability investigations

    Directory of Open Access Journals (Sweden)

    Spinks CB

    2017-02-01

    Full Text Available Crystal B Spinks,1 Ahmed S Zidan,2,3 Mansoor A Khan,4 Muhammad J Habib,1 Patrick J Faustino2 1Department of Pharmaceutical Sciences, School of Pharmacy, Howard University, Washington, DC, 2Division of Product Quality Research, Office of Pharmaceutical Quality, Food and Drug Administration, Silver Spring, MD, USA; 3Faculty of Pharmacy, Zagazig University, Zagazig, Egypt; 4Irma Lerma Rangel College of Pharmacy, Texas A&M Health Science Center, College Station, TX, USA Abstract: Tenofovir, currently marketed as the prodrug tenofovir disoproxil fumarate, is used clinically to treat patients with HIV/AIDS. The oral bioavailability of tenofovir is relatively low, limiting its clinical effectiveness. Encapsulation of tenofovir within modified long-circulating liposomes would deliver this hydrophilic anti-HIV drug to the reticuloendothelial system for better therapeutic efficacy. The objectives of the current study were to prepare and pharmaceutically characterize model liposomal tenofovir formulations in an attempt to improve their bioavailability. The entrapment process was performed using film hydration method, and the formulations were characterized in terms of encapsulation efficiency and Caco-2 permeability. An efficient reverse-phase high-performance liquid chromatography method was developed and validated for tenofovir quantitation in both in vitro liposomal formulations and Caco-2 permeability samples. Separation was achieved isocratically on a Waters Symmetry C8 column using 10 mM Na2PO4/acetonitrile pH 7.4 (95:5 v/v. The flow rate was 1 mL/min with a 12 min elution time. Injection volume was 10 µL with ultraviolet detection at 270 nm. The method was validated according to United States Pharmacopeial Convention category I requirements. The obtained result showed that tenofovir encapsulation within the prepared liposomes was dependent on the employed amount of the positive charge-imparting agent. The obtained results indicated that

  17. Pilot Investigation of 2 Nondiet Approaches to Improve Weight and Health.

    Science.gov (United States)

    Webber, Kelly H; Mellin, Laurel; Mayes, Lindsey; Mitrovic, Igor; Saulnier, Matt

    2017-06-23

    be viable as an approach to weight loss and improvements in health-related outcomes in the short term. A longer investigation of the program is warranted.

  18. Combined approach based on principal component analysis and canonical discriminant analysis for investigating hyperspectral plant response

    Directory of Open Access Journals (Sweden)

    Anna Maria Stellacci

    2012-07-01

    Full Text Available Hyperspectral (HS data represents an extremely powerful means for rapidly detecting crop stress and then aiding in the rational management of natural resources in agriculture. However, large volume of data poses a challenge for data processing and extracting crucial information. Multivariate statistical techniques can play a key role in the analysis of HS data, as they may allow to both eliminate redundant information and identify synthetic indices which maximize differences among levels of stress. In this paper we propose an integrated approach, based on the combined use of Principal Component Analysis (PCA and Canonical Discriminant Analysis (CDA, to investigate HS plant response and discriminate plant status. The approach was preliminary evaluated on a data set collected on durum wheat plants grown under different nitrogen (N stress levels. Hyperspectral measurements were performed at anthesis through a high resolution field spectroradiometer, ASD FieldSpec HandHeld, covering the 325-1075 nm region. Reflectance data were first restricted to the interval 510-1000 nm and then divided into five bands of the electromagnetic spectrum [green: 510-580 nm; yellow: 581-630 nm; red: 631-690 nm; red-edge: 705-770 nm; near-infrared (NIR: 771-1000 nm]. PCA was applied to each spectral interval. CDA was performed on the extracted components to identify the factors maximizing the differences among plants fertilised with increasing N rates. Within the intervals of green, yellow and red only the first principal component (PC had an eigenvalue greater than 1 and explained more than 95% of total variance; within the ranges of red-edge and NIR, the first two PCs had an eigenvalue higher than 1. Two canonical variables explained cumulatively more than 81% of total variance and the first was able to discriminate wheat plants differently fertilised, as confirmed also by the significant correlation with aboveground biomass and grain yield parameters. The combined

  19. Investigation into in vitro anti-leishmanial combinations of calcium channel blockers and current anti-leishmanial drugs

    Directory of Open Access Journals (Sweden)

    Juliana Quero Reimão

    2011-12-01

    Full Text Available The need for drug combinations to treat visceral leishmaniasis (VL arose because of resistance to antimonials, the toxicity of current treatments and the length of the course of therapy. Calcium channel blockers (CCBs have shown anti-leishmanial activity; therefore their use in combination with standard drugs could provide new alternatives for the treatment of VL. In this work, in vitro isobolograms of Leishmania (Leishmania chagasi using promastigotes or intracellular amastigotes were utilised to identify the interactions between five CCBs and the standard drugs pentamidine, amphotericin B and glucantime. The drug interactions were assessed with a fixed ratio isobologram method and the fractional inhibitory concentrations (FICs, sum of FICs (ΣFICs and the overall mean ΣFIC were calculated for each combination. Graphical isobologram analysis showed that the combination of nimodipine and glucantime was the most promising in amastigotes with an overall mean ΣFIC value of 0.79. Interactions between CCBs and the anti-leishmanial drugs were classified as indifferent according to the overall mean ΣFIC and the isobologram graphic analysis.

  20. In vitro and in vivo investigation of bacterial cellulose dressing containing uniform silver sulfadiazine nanoparticles for burn wound healing

    Directory of Open Access Journals (Sweden)

    Xiaoxiao Wen

    2015-06-01

    Full Text Available Silver sulfadiazine (SSD particles in homogeneous dispersion state were prepared by an ultrasonic method and then nano- and microparticles were separated using centrifugation. SSD particles with narrow size distribution were impregnated with bacterial cellulose (BC to produce BC–SSD composite membrane used as burn wound dressing. A scanning electron microscope (SEM was used to examine the surface morphology of BC–SSD membranes. The incorporation of SSD in BC–SSD was confirmed by X-ray diffraction (XRD. Antimicrobial tests in vitro indicated that BC–SSD showed excellent antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. The effects of BC–SSD on burn wound healing were assessed by rat models. The comparative study confirmed that the wound treated with BC–SSD showed high healing rate. The bacteria count in BC–SSD group was far less than control group. Histological analysis showed that epithelialization progressed better in wound treated with BC–SSD. These values demonstrated that the BC–SSD composite membrane could be a promising wound dressing for burn.

  1. Transfer of uranium throughout the entire gastrointestinal tract of the rat: In vivo and in vitro approaches

    Energy Technology Data Exchange (ETDEWEB)

    Dublineau, I.; Grison, S.; Dudoignon, N.; Baudelin, C.; Aigueperse, J.

    2004-07-01

    The presence of uranium in environment either natural or due to civil and military use, may lead to contamination of the public throughout the entire life mainly by chronic ingestion. The mechanisms of uranium transfer from alimentary bolus to blood are still not well known. In particular, few information are available on the different absorption sites along the gastrointestinal tract, the different cellular pathways (para-or trans-cellular), and the transporters implicated in the uranium absorption. In addition, the specific role of Peyer's patches, the aggregated structure of Gut-Associated Lymphoid tissue, in the intestinal transfer of uranium has never been determined. In fact, the transport of uranium through these structures specialized in antigen uptake from intestinal lumen may lead to major dysfunctions in mucosal immunity. Thus, different approaches have to be developed to determine the role of the different gastrointestinal structures and to apprehend the biological consequences of daily passage of uranium through these structures. These experiments include in vivo measurement of uranium in blood after in situ deposit of uranium (233U) in the different segments of the alimentary tract (buccal cavity, stomach, small intestine, colon) and ex vivo experiments in Using chambers to compare uranium passage from luminal to serosal side through intestinal epithelium and Peyer's patches. In vitro studies are also necessary to determine the nature of the cells as well as the transporters implicated in the gastrointestinal passage of uranium. Autoradiography experiments were performed to determine if uranium absorption was only restricted to villi which contained absorptive cells or if uranium absorption was also due to crypt cells. In addition, the transporter implicated in the uranium passage is dependent of the physico-chemical form of uranium present at the different gastrointestinal sites. When complexed to phosphate, uranium is transported by the

  2. Immunogenicity investigations of lipidoid structures in vitro and in silico: Modulating lipidoid-mediated TLR4 activation by nanoparticle design

    DEFF Research Database (Denmark)

    de Groot, Anne Marit; Thanki, Kaushik; Gangloff, Monique

    2018-01-01

    Therapeutics based on small interfering RNA (siRNA) have promising potential as antiviral and anti-inflammatory agents. To deliver siRNA across cell membranes to reach the RNA interference pathway in the cytosol of target cells, non-viral nanoparticulate delivery approaches are explored. Recently...

  3. Investigation of geometrical effects in the carbon allotropes manipulation based on AFM: multiscale approach

    Energy Technology Data Exchange (ETDEWEB)

    Korayem, M. H., E-mail: hkorayem@iust.ac.ir; Hefzabad, R. N.; Homayooni, A.; Aslani, H. [Iran University of Science and Technology, Robotic Research Laboratory, Center of Excellence in Experimental Solid Mechanics and Dynamics, School of Mechanical Engineering (Iran, Islamic Republic of)

    2017-01-15

    Carbon allotropes are used as nanocarriers for drug and cell delivery. To obtain an accurate result in the nanoscale, it is important to use a precise model. In this paper, a multiscale approach is presented to investigate the manipulation process of carbon allotropes based on atomic force microscopy (AFM). For this purpose, the AFM setup is separated into two parts with different sizes as macro field (MF) and nano field (NF). Using Kirchhoff’s plate model, the cantilever (the main part of MF) is modeled. The molecular dynamics method is applied to model the NF part, and then the MF and NF are coupled with the multiscale algorithm. With this model, by considering the effect of size and shape, the manipulation of carbon allotropes is carried out. The manipulations of armchair CNTs and fullerenes are performed to study the diameter changing effects. The result shows that the manipulation and friction force increases by increasing the diameter. The result of the indentation depth for the armchair CNTs indicates that decreasing the diameter causes the indentation depth to reduce. Moreover, the manipulations of four kinds of carbon allotropes with the same number of atoms have been studied to investigate the geometrical effects. The shapes of these nanoparticles change from sphere to cylinder. The results illustrate that the manipulation and the friction force decrease as the nanoparticle shape varies from sphere to cylinder. The Von-Mises results demonstrate that by changing the nanoparticle shape from the spherical to the cylindrical form, the stress increases, although the manipulation force reduces.

  4. Field Investigation of a New Recharge Approach for ASR Projects in Near-Surface Aquifers.

    Science.gov (United States)

    Liu, Gaisheng; Knobbe, Steven; Reboulet, Edward C; Whittemore, Donald O; Händel, Falk; Butler, James J

    2016-05-01

    Aquifer storage and recovery (ASR) is the artificial recharge and temporary storage of water in an aquifer when water is abundant, and recovery of all or a portion of that water when it is needed. One key limiting factor that still hinders the effectiveness of ASR is the high costs of constructing, maintaining, and operating the artificial recharge systems. Here we investigate a new recharge method for ASR in near-surface unconsolidated aquifers that uses small-diameter, low-cost wells installed with direct-push (DP) technology. The effectiveness of a DP well for ASR recharge is compared with that of a surface infiltration basin at a field site in north-central Kansas. The performance of the surface basin was poor at the site due to the presence of a shallow continuous clay layer, identified with DP profiling methods, that constrained the downward movement of infiltrated water and significantly reduced the basin recharge capacity. The DP well penetrated through this clay layer and was able to recharge water by gravity alone at a much higher rate. Most importantly, the costs of the DP well, including both the construction and land costs, were only a small fraction of those for the infiltration basin. This low-cost approach could significantly expand the applicability of ASR as a water resources management tool to entities with limited fiscal resources, such as many small municipalities and rural communities. The results of this investigation demonstrate the great potential of DP wells as a new recharge option for ASR projects in near-surface unconsolidated aquifers. © 2015, National Ground Water Association.

  5. ConMap: Investigating new computer-based approaches to assessing conceptual knowledge structure in physics

    Science.gov (United States)

    Beatty, Ian D.

    2000-06-01

    There is a growing consensus among educational researchers that traditional problem-based assessments are not effective tools for diagnosing a student's knowledge state and for guiding pedagogical intervention, and that new tools grounded in the results of cognitive science research are needed. The ConMap (``Conceptual Mapping'') project, described in this dissertation, proposed and investigated some novel methods for assessing the conceptual knowledge structure of physics students. A set of brief computer-administered tasks for eliciting students' conceptual associations was designed. The basic approach of the tasks was to elicit spontaneous term associations from subjects by presenting them with a prompt term, or problem, or topic area, and having them type a set of response terms. Each response was recorded along with the time spent thinking of and typing it. Several studies were conducted in which data was collected on introductory physics students' performance on the tasks. A detailed statistical description of the data was compiled. Phenomenological characterization of the data (description and statistical summary of observed patterns) provided insight into the way students respond to the tasks, and discovered some notable features to guide modeling efforts. Possible correlations were investigated, some among different aspects of the ConMap data, others between aspects of the data and students' in-course exam scores. Several correlations were found which suggest that the ConMap tasks can successfully reveal information about students' knowledge structuring and level of expertise. Similarity was observed between data from one of the tasks and results from a traditional concept map task. Two rudimentary quantitative models for the temporal aspects of student performance on one of the tasks were constructed, one based on random probability distributions and the other on a detailed deterministic representation of conceptual knowledge structure. Both models were

  6. Interactions among biotic and abiotic factors affect the reliability of tungsten microneedles puncturing in vitro and in vivo peripheral nerves: A hybrid computational approach.

    Science.gov (United States)

    Sergi, Pier Nicola; Jensen, Winnie; Yoshida, Ken

    2016-02-01

    Tungsten is an elective material to produce slender and stiff microneedles able to enter soft tissues and minimize puncture wounds. In particular, tungsten microneedles are used to puncture peripheral nerves and insert neural interfaces, bridging the gap between the nervous system and robotic devices (e.g., hand prostheses). Unfortunately, microneedles fail during the puncture process and this failure is not dependent on stiffness or fracture toughness of the constituent material. In addition, the microneedles' performances decrease during in vivo trials with respect to the in vitro ones. This further effect is independent on internal biotic effects, while it seems to be related to external biotic causes. Since the exact synergy of phenomena decreasing the in vivo reliability is still not known, this work explored the connection between in vitro and in vivo behavior of tungsten microneedles through the study of interactions between biotic and abiotic factors. A hybrid computational approach, simultaneously using theoretical relationships and in silico models of nerves, was implemented to model the change of reliability varying the microneedle diameter, and to predict in vivo performances by using in vitro reliability and local differences between in vivo and in vitro mechanical response of nerves. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. An investigation of the relationship between the anti-inflammatory activity, polyphenolic content, and antioxidant activities of cooked and in vitro digested culinary herbs.

    Science.gov (United States)

    Chohan, Magali; Naughton, Declan P; Jones, Lucy; Opara, Elizabeth I

    2012-01-01

    There is little research on how cooking and digestion affect the anti-inflammatory activity of culinary herbs. Thus, the aim of this paper was to investigate this activity following cooking and in vitro digestion of the common culinary herbs, rosemary, sage, and thyme, and the relationship between their anti-inflammatory activity, polyphenol content, and antioxidant capacity. The anti-inflammatory activity of uncooked (U), cooked (C), cooked and in vitro digested (C&D), and standardised (STD, 30 mg/mL) culinary herbs was assessed by measuring their effect on interleukin 8 (IL-8) release from stimulated human peripheral blood lymphocytes (PBLs) and Caco-2 cells. The trolox equivalent capacity (TEAC) and estimated total phenolic content of the herbs were also determined. There was a significant decrease in IL-8 release from PBLs stimulated with H(2)O(2) incubated with (U), (C), (C&D), and (STD) herbs and from Caco-2 cells stimulated with TNFα incubated with (C&D) and (STD) herbs. PBLs pre-incubated with (C&D) herbs prior to stimulation (H(2)O(2) or TNFα) caused a significant inhibition in IL-8 release. The significant correlations between TEAC and estimated phenolic content and the anti-inflammatory activity suggest a possible contributory role of polyphenols to the anti-inflammatory activity of the culinary herbs investigated.

  8. INVESTIGATIONS ON A COMBINED RGB / TIME-OF-FLIGHT APPROACH FOR CLOSE RANGE APPLICATIONS

    Directory of Open Access Journals (Sweden)

    H. Hastedt

    2012-07-01

    Full Text Available 3D surface and scene reconstruction for close range applications mainly rely on high resolution and accurate system devices and powerful algorithms. Camera systems based on the time-of-flight principle allow for real-time 3D distance measurements. Unfortunately these devices are limited in resolution and accuracy. But applying calibration models and combining with high-resolution image data offers a promising approach in order to form a multisensor system for close range applications. This article will present investigations on such a multisensor system. Different options on data fusion processing of distance information and high-resolution color information in order to generate dense 2 1/2 D and 3D point clouds will be presented. The multisensor system is calibrated with respect to its interior and exterior orientation. The time-of-flight distance information is optimized extracting best information of different data captures with a set of integration times following the principle of high dynamic range imaging. The high-resolution RGB image information is projected into object space and intersected with the object surface from the time-of-flight camera. First results of this solution on dense monoplotting and its verification will be presented.

  9. [Investigation of the sudden infant death syndrome: a multidisciplinary approach is required].

    Science.gov (United States)

    Limelette, Anne; Boulagnon, Camille; Terrade, Caroline; N'guyen, Yohan; Guillard, Thomas; Andréoletti, Laurent; Garnotel, Roselyne; Digeon, Béatrice; De Champs, Christophe; Fornes, Paul; Lévêque, Nicolas

    2013-01-01

    The concept of sudden infant death syndrome (SIDS) is defined as the sudden, unexpected death of an infant less than a year old which remains unexplained after in-depth investigations comprising a complete autopsy, biological analyses, and a clinical examination of the circumstances surrounding the death. This definition underlines the importance of finding the cause of this disease in order to improve preventative measures to reduce the number of deaths due to sudden infant death syndrome. Among the causes of SIDS, pediatric infectious diseases may be neglected and must be systematically sought after. We report upon a SIDS death case of a four and a half month-old that occurred during his sleep. Following the absence of an evident cause of death a scientific autopsy was performed. The histological examination of pulmonary tissue revealed broncolitic lesions associated with numerous micro-abscesses. The post mortem microbiological analyses revealed evidence of an infection by the respiratory syncytial virus complicated by a bacterial infection due to Haemophilus influenzae. The case underlines the necessity of a multidisciplinary approach to researching SIDS, involving both clinicians and biologists, in order to determine the causes of these deaths.

  10. Computational investigation of fluid flow and heat transfer of an economizer by porous medium approach

    Science.gov (United States)

    Babu, C. Rajesh; Kumar, P.; Rajamohan, G.

    2017-07-01

    Computation of fluid flow and heat transfer in an economizer is simulated by a porous medium approach, with plain tubes having a horizontal in-line arrangement and cross flow arrangement in a coal-fired thermal power plant. The economizer is a thermal mechanical device that captures waste heat from the thermal exhaust flue gasses through heat transfer surfaces to preheat boiler feed water. In order to evaluate the fluid flow and heat transfer on tubes, a numerical analysis on heat transfer performance is carried out on an 110 t/h MCR (Maximum continuous rating) boiler unit. In this study, thermal performance is investigated using the computational fluid dynamics (CFD) simulation using ANSYS FLUENT. The fouling factor ε and the overall heat transfer coefficient ψ are employed to evaluate the fluid flow and heat transfer. The model demands significant computational details for geometric modeling, grid generation, and numerical calculations to evaluate the thermal performance of an economizer. The simulation results show that the overall heat transfer coefficient 37.76 W/(m2K) and economizer coil side pressure drop of 0.2 (kg/cm2) are found to be conformity within the tolerable limits when compared with existing industrial economizer data.

  11. Investigation on the relationship between solubility of artemisinin and polyvinylpyrroli done addition by using DAOSD approach.

    Science.gov (United States)

    Zhang, Jin; Guo, Ran; He, Anqi; Weng, Shifu; Gao, Xiuxiang; Xu, Yizhuang; Noda, Isao; Wu, Jinguang

    2017-07-05

    In this work, we investigated the influence of polyvinylpyrrolidone (PVP) on the solubility of artemisinin in aqueous solution by using quantitative 1H NMR. Experimental results demonstrate that about 4 times of incremental increase occurs on the solubility of artemisinin upon introducing PVP. In addition, dipole-dipole interaction between the ester group of artemisinin and the amide group of N-methylpyrrolidone (NMP), a model compound of PVP, is characterized by two-dimensional (2D) correlation FTIR spectroscopy with the DAOSD (Double Asynchronous Orthogonal Sample Design) approach developed in our previous work. The observation of cross peaks in a pair of 2D asynchronous spectra suggests that dipole-dipole interaction indeed occurs between the ester group of artemisinin and amide group of NMP. Moreover, the pattern of cross peaks indicates that the carbonyl band of artemisinin undergoes blue-shift while the bandwidth and absorptivity increases via interaction with NMP, and the amide band of NMP undergoes blue-shift while the absorptivity increases via interaction with artemisinin. Dipole-dipole interaction, as one of the strongest intermolecular interaction between artemisinin and excipient, may play an important role in the enhancement of the solubility of artemisinin in aqueous solution. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Validation of a general in vitro approach for prediction of total lung deposition in healthy adults for pharmaceutical inhalation products.

    Science.gov (United States)

    Olsson, Bo; Borgström, Lars; Lundbäck, Hans; Svensson, Mårten

    2013-12-01

    A validated method to predict lung deposition for inhaled medication from in vitro data is lacking in spite of many attempts to correlate in vitro and in vivo outcomes. By using an in vivo-like in vitro setup and analyzing inhalers from the same batches, both in vitro and in vivo, we wanted to create a situation where information from the in vitro and in vivo outcomes could be analyzed at the same time. Nine inhalation products containing either budesonide or AZD4818 were evaluated. These comprised two pressurized metered dose inhalers (pMDIs), a pMDI plus a spacer, four dry powder inhalers, and two dosimetric nebulizers. In vitro, an in vivo-like setup consisting of anatomically correct inlet throats were linked to a flow system that could replay actual inhalation flow profiles through the throat to a filter or to an impactor. In vivo, total lung deposition was measured in healthy adults by pharmacokinetic methods. We could show that the amount of drug escaping filtration in a realistic throat model under realistic delivery conditions predicts the typical total lung deposition in trained healthy adult subjects in the absence of significant exhaled mass. We could further show that by using combinations of throat models and flow profiles that represent realistic deviations from the typical case, variations in ex-cast deposition reflect between-subject variation in lung deposition. Further, we have demonstrated that ex-cast deposition collected either by a simple filter or by a cascade impactor operated at a fixed flow rate using a mixing inlet, to accommodate a variable flow profile through the inhaler, predicts equally well the lung deposited dose. Additionally, the ex-cast particle size distribution measured by this method may be relevant for predicting exhaled fraction and regional lung deposition by computational models.

  13. On the potential of a new IVUS elasticity modulus imaging approach for detecting vulnerable atherosclerotic coronary plaques: in vitro vessel phantom study.

    Science.gov (United States)

    Le Floc'h, Simon; Cloutier, Guy; Finet, Gérard; Tracqui, Philippe; Pettigrew, Roderic I; Ohayon, Jacques

    2010-10-07

    Peak cap stress amplitude is recognized as a good indicator of vulnerable plaque (VP) rupture. However, such stress evaluation strongly relies on a precise, but still lacking, knowledge of the mechanical properties exhibited by the plaque components. As a first response to this limitation, our group recently developed, in a previous theoretical study, an original approach, called iMOD (imaging modulography), which reconstructs elasticity maps (or modulograms) of atheroma plaques from the estimation of strain fields. In the present in vitro experimental study, conducted on polyvinyl alcohol cryogel arterial phantoms, we investigate the benefit of coupling the iMOD procedure with the acquisition of intravascular ultrasound (IVUS) measurements for detection of VP. Our results show that the combined iMOD-IVUS strategy: (1) successfully detected and quantified soft inclusion contours with high positive predictive and sensitivity values of 89.7 ± 3.9% and 81.5 ± 8.8%, respectively, (2) estimated reasonably cap thicknesses larger than ∼300 µm, but underestimated thinner caps, and (3) quantified satisfactorily Young's modulus of hard medium (mean value of 109.7 ± 23.7 kPa instead of 145.4 ± 31.8 kPa), but overestimated the stiffness of soft inclusions (mean Young`s moduli of 31.4 ± 9.7 kPa instead of 17.6 ± 3.4 kPa). All together, these results demonstrate a promising benefit of the new iMOD-IVUS clinical imaging method for in vivo VP detection.

  14. On the potential of a new IVUS elasticity modulus imaging approach for detecting vulnerable atherosclerotic coronary plaques: in vitro vessel phantom study

    Energy Technology Data Exchange (ETDEWEB)

    Floc' h, Simon Le; Tracqui, Philippe; Ohayon, Jacques [Laboratory TIMC-DynaCell, UJF, CNRS UMR 5525, In3S, Grenoble (France); Cloutier, Guy [Laboratory of Biorheology and Medical Ultrasonics, Research Center, University of Montreal Hospital (CRCHUM), Montreal, Quebec (Canada); Finet, Gerard [Department of Hemodynamics and Interventional Cardiology, Hospices Civils de Lyon and Claude Bernard University Lyon 1, INSERM Unit 886, Lyon (France); Pettigrew, Roderic I, E-mail: Guy.Cloutier@umontreal.c, E-mail: Jacques.Ohayon@imag.f [Laboratory of Integrative Cardiovascular Imaging Science, National Institute of Diabetes Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD (United States)

    2010-10-07

    Peak cap stress amplitude is recognized as a good indicator of vulnerable plaque (VP) rupture. However, such stress evaluation strongly relies on a precise, but still lacking, knowledge of the mechanical properties exhibited by the plaque components. As a first response to this limitation, our group recently developed, in a previous theoretical study, an original approach, called iMOD (imaging modulography), which reconstructs elasticity maps (or modulograms) of atheroma plaques from the estimation of strain fields. In the present in vitro experimental study, conducted on polyvinyl alcohol cryogel arterial phantoms, we investigate the benefit of coupling the iMOD procedure with the acquisition of intravascular ultrasound (IVUS) measurements for detection of VP. Our results show that the combined iMOD-IVUS strategy: (1) successfully detected and quantified soft inclusion contours with high positive predictive and sensitivity values of 89.7 {+-} 3.9% and 81.5 {+-} 8.8%, respectively, (2) estimated reasonably cap thicknesses larger than {approx}300 {mu}m, but underestimated thinner caps, and (3) quantified satisfactorily Young's modulus of hard medium (mean value of 109.7 {+-} 23.7 kPa instead of 145.4 {+-} 31.8 kPa), but overestimated the stiffness of soft inclusions (mean Young's moduli of 31.4 {+-} 9.7 kPa instead of 17.6 {+-} 3.4 kPa). All together, these results demonstrate a promising benefit of the new iMOD-IVUS clinical imaging method for in vivo VP detection.

  15. Systems biology approach to transplant tolerance: proof of concept experiments using RNA interference (RNAi) to knock down hub genes in Jurkat and HeLa cells in vitro.

    Science.gov (United States)

    Lwin, Wint Wah; Park, Ken; Wauson, Matthew; Gao, Qin; Finn, Patricia W; Perkins, David; Khanna, Ajai

    2012-07-01

    Systems biology is gaining importance in studying complex systems such as the functional interconnections of human genes [1]. To investigate the molecular interactions involved in T cell immune responses, we used databases of physical gene-gene interactions to constructed molecular interaction networks (interconnections) with R language algorithms. This helped to identify highly interconnected "hub" genes AT(1)P5C1, IL6ST, PRKCZ, MYC, FOS, JUN, and MAPK1. We hypothesized that suppression of these hub genes in the gene network would result in significant phenotypic effects on T cells and examined this in vitro. The molecular interaction networks were then analyzed and visualized with Cytoscape. Jurkat and HeLa cells were transfected with siRNA for the selected hub genes. Cell proliferation was measured using ATP luminescence and BrdU labeling, which were measured 36, 72, and 96 h after activation. Following T cell stimulation, we found a significant decrease in ATP production (P cells. However, HeLa cells showed a significant (P cell proliferation when the genes MAPK1, IL6ST, ATP5C1, JUN, and FOS were knocked down. In both Jurkat and HeLa cells, targeted gene knockdown using siRNA showed decreased cell proliferation and ATP production in both Jurkat and HeLa cells. However, Jurkat T cells and HELA cells use different hub genes to regulate activation responses. This experiment provides proof of principle of applying siRNA knockdown of T cell hub genes to evaluate their proliferative capacity and ATP production. This novel concept outlines a systems biology approach to identify hub genes for targeted therapeutics. Published by Elsevier Inc.

  16. In vitro investigation on extensively destroyed vital teeth: is fracture force a limiting factor for direct restoration?

    Science.gov (United States)

    Schwindling, F S; Hartmann, T; Panagidis, D; Krisam, J; Rues, S; Schmitter, M

    2014-12-01

    To evaluate the in vitro fracture load of extensively damaged vital teeth after either direct or indirect restauration, severe tooth substance loss was simulated for 96 molars. Subsequently, two cavities were prepared with little (design 1) or more substantial (design 2) residual tooth support. All molars were provided with a 2-mm ferrule design and then divided into 12 test groups based on their occlusal surface size. They were restored with composite or with either of two types of single crown (cast metal or milled zirconia). After thermal ageing (10,000 cycles at 6.5 and 60 °C), 1.2 million cycles of chewing simulation were applied (64 N). Maximum fracture load was determined with a loading angle of 45°. Statistical analysis was performed by use of Kaplan-Meier modelling, Student's t-tests, one-way anova, post hoc Tukey's HSD tests and linear regression analysis. Regarding mean fracture load without ageing, the indirect restorations outperformed composite (design 1: direct: 508 ± 123 N, indirect: 741 ± 248 N; design 2: direct: 554 ± 167 N, indirect: 903 ± 221 N). After artificial ageing, however, these differences were no longer significant (design 1: direct: 328 ± 189 N, indirect: 506 ± 352 N; design 2: direct 399 ± 208 N, indirect 577 ± 292 N). Instead, the fracture load of the aged composite restorations was comparable with that for zirconia (design 1) and cast metal (design 2) crowns. Fracture loads of direct composite restorations after artificial ageing might fulfil clinical requirements. © 2014 John Wiley & Sons Ltd.

  17. Neuronal hypoxia in vitro: Investigation of therapeutic principles of HUCB-MNC and CD133+ stem cells

    Directory of Open Access Journals (Sweden)

    Emmrich Frank

    2008-09-01

    Full Text Available Abstract Background The therapeutic capacity of human umbilical cord blood mononuclear cells (HUCB-MNC and stem cells derived thereof is documented in animal models of focal cerebral ischemia, while mechanisms behind the reduction of lesion size and the observed improvement of behavioral skills still remain poorly understood. Methods A human in vitro model of neuronal hypoxia was used to address the impact of total HUCB-MNC (tMNC, a stem cell enriched fraction (CD133+, 97.38% CD133-positive cells and a stem cell depleted fraction (CD133-, 0.06% CD133-positive cells of HUCB-MNC by either direct or indirect co-cultivation with post-hypoxic neuronal cells (differentiated SH-SY5Y. Over three days, development of apoptosis and necrosis of neuronal cells, chemotaxis of MNC and production of chemokines (CCL2, CCL3, CCL5, CXCL8, CXCL9 and growth factors (G-CSF, GM-CSF, VEGF, bFGF were analyzed using fluorescence microscopy, FACS and cytometric bead array. Results tMNC, CD133+ and surprisingly CD133- reduced neuronal apoptosis in direct co-cultivations significantly to levels in the range of normoxic controls (7% ± 3%. Untreated post-hypoxic control cultures showed apoptosis rates of 85% ± 11%. tMNC actively migrated towards injured neuronal cells. Both co-cultivation types using tMNC or CD133- reduced apoptosis comparably. CD133- produced high concentrations of CCL3 and neuroprotective G-CSF within indirect co-cultures. Soluble factors produced by CD133+ cells were not detectable in direct co-cultures. Conclusion Our data show that heterogeneous tMNC and even CD133-depleted fractions have the capability not only to reduce apoptosis in neuronal cells but also to trigger the retaining of neuronal phenotypes.

  18. Assessing the risk of alcohol-induced dose dumping from sustained-release oral dosage forms: in vitro-in silico approach.

    Science.gov (United States)

    Cvijić, Sandra; Aleksić, Ivana; Ibrić, Svetlana; Parojčić, Jelena

    2017-10-29

    Consumption of alcoholic beverages with sustained-release oral dosage forms may pose a risk to patients due to potential alcohol-induced dose dumping (ADD). Regulatory guidances recommend in vitro dissolution testing to identify the risk of ADD, but the question remains whether currently proposed test conditions can be considered biopredictive. The purpose of this study was to evaluate different dissolution setups to assess ADD, and the potential of combined in vitro-in silico approach to predict drug absorption after concomitant alcohol intake for hydrophilic and lipophilic sustained-release tablets containing ibuprofen or diclofenac sodium. According to the obtained results, the impact of ethanol was predominantly governed by the influence on matrix integrity, with the increase in drug solubility being less significant. Hydrophilic matrix tablets were less susceptible to ADD than lipophilic matrices, although the conclusion on formulation ethanol-vulnerability depended on the employed experimental conditions. In silico predictions indicated that the observed changes in drug dissolution would not result in plasma concentrations beyond therapeutic window, but sustained-release characteristics of the formulations might be lost. Overall, the study demonstrated that in vitro-in silico approach may provide insight into the effect of ADD on drug clinical performance, and serve as a tool for ADD risk assessment.

  19. Integrated in vitro approaches to assess the bioaccessibility and bioavailability of silicon-biofortified leafy vegetables and preliminary effects on bone.

    Science.gov (United States)

    D'Imperio, Massimiliano; Brunetti, Giacomina; Gigante, Isabella; Serio, Francesco; Santamaria, Pietro; Cardinali, Angela; Colucci, Silvia; Minervini, Fiorenza

    2017-03-01

    Food industries are increasingly oriented toward new foods to improve nutritional status and/or to combat nutritional deficiency diseases. In this context, silicon biofortification could be an innovative tool for obtaining new foods with possible positive effects on bone mineralization. In this paper, an alternative and quick in vitro approach was applied in order to evaluate the potential health-promoting effects of five silicon-biofortified leafy vegetables (tatsoi, mizuna, purslane, Swiss chard and chicory) on bone mineralization compared with a commercial silicon supplement. The silicon bioaccessibility and bioavailability of the five leafy vegetables (biofortified or not) and of the supplement were assessed by applying a protocol consisting of in vitro gastrointestinal digestion coupled with a Caco-2 cell model. Silicon bioaccessibility ranged from 0.89 to 8.18 mg/L and bioavailability ranged from 111 to 206 μg/L of Si for both vegetables and supplement. Furthermore, the bioavailable fractions were tested on a human osteoblast cell model following the expression of type 1 collagen and alkaline phosphatase. The results obtained highlighted that the bioavailable fraction of biofortified purslane and Swiss chard improved the expression of both osteoblast markers compared with the supplement and other vegetables. These results underline the potentially beneficial effect of biofortified leafy vegetables and also indicate the usefulness of in vitro approaches for selecting the best vegetable with positive bone effects for further in vivo research.

  20. 76 FR 53683 - Draft Guidance for Industry on Oversight of Clinical Investigations: A Risk-Based Approach to...

    Science.gov (United States)

    2011-08-29

    ... HUMAN SERVICES Food and Drug Administration Draft Guidance for Industry on Oversight of Clinical... guidance for industry entitled ``Oversight of Clinical Investigations: A Risk-Based Approach to Monitoring... for clinical investigations of human drugs, biologics, medical devices, and combinations thereof. The...

  1. 78 FR 48173 - Guidance for Industry on Oversight of Clinical Investigations-A Risk-Based Approach to Monitoring...

    Science.gov (United States)

    2013-08-07

    ... No. FDA-2011-D-0597] Guidance for Industry on Oversight of Clinical Investigations--A Risk-Based... entitled ``Oversight of Clinical Investigations--A Risk-Based Approach to Monitoring.'' This guidance... human drugs, biologics, medical devices, and combinations thereof. The overarching goal of this guidance...

  2. Development of a Physiologically Relevant Population Pharmacokinetic in Vitro-in Vivo Correlation Approach for Designing Extended-Release Oral Dosage Formulation.

    Science.gov (United States)

    Kim, Tae Hwan; Shin, Soyoung; Bulitta, Jürgen B; Youn, Yu Seok; Yoo, Sun Dong; Shin, Beom Soo

    2017-01-03

    Establishing a level A in vitro-in vivo correlation (IVIVC) for a drug with complex absorption kinetics is challenging. The objective of the present study was to develop an IVIVC approach based on population pharmacokinetic (POP-PK) modeling that incorporated physiologically relevant absorption kinetics. To prepare three extended release (ER) tablets of loxoprofen, three types of hydroxypropyl methylcellulose (HPMC 100, 4000, and 15000 cps) were used as drug release modifiers, while lactose and magnesium stearate were used as the diluent and lubricant, respectively. An in vitro dissolution test in various pH conditions showed that loxoprofen dissolution was faster at higher pH. The in vivo pharmacokinetics of loxoprofen was assessed following oral administration of the different loxoprofen formulations to Beagle dogs (n = 22 in total). Secondary peaks or shoulders were observed in many of the individual plasma concentration vs time profiles after ER tablet administration, which may result from secondary absorption in the intestine due to a dissolution rate increase under intestinal pH compared to that observed at stomach pH. In addition, in vivo oral bioavailability was found to decrease with prolonged drug dissolution, indicating site-specific absorption. Based on the in vitro dissolution and in vivo absorption data, a POP-PK IVIVC model was developed using S-ADAPT software. pH-dependent biphasic dissolution kinetics, described using modified Michaelis-Menten kinetics with varying Vmax, and site-specific absorption, modeled using a changeable absorbed fraction parameter, were applied to the POP-PK IVIVC model. To experimentally determine the biphasic dissolution profiles of the ER tablets, another in vitro dissolution test was conducted by switching dissolution medium pH based on an in vivo estimate of gastric emptying time. The model estimated, using linear regression, that in vivo initial maximum dissolution rate (Vmax(0)in vivo) was highly correlated (r2 > 0

  3. Porcine oocyte maturation in vitro: role of cAMP and oocyte-secreted factors – A practical approach

    Science.gov (United States)

    APPELTANT, Ruth; SOMFAI, Tamás; MAES, Dominiek; VAN SOOM, Ann; KIKUCHI, Kazuhiro

    2016-01-01

    Polyspermy or the penetration of more than one sperm cell remains a problem during porcine in vitro fertilization (IVF). After in vitro culture of porcine zygotes, only a low percentage of blastocysts develop and their quality is inferior to that of in vivo derived blastocysts. It is unknown whether the cytoplasmic maturation of the oocyte is sufficiently sustained in current in vitro maturation (IVM) procedures. The complex interplay between oocyte and cumulus cells during IVM is a key factor in this process. By focusing on this bidirectional communication, it is possible to control the coordination of cumulus expansion, and nuclear and cytoplasmic maturation during IVM to some extent. Therefore, this review focuses on the regulatory mechanisms between oocytes and cumulus cells to further the development of new in vitro embryo production (IVP) procedures, resulting in less polyspermy and improved oocyte developmental potential. Specifically, we focused on the involvement of cAMP in maturation regulation and function of oocyte-secreted factors (OSFs) in the bidirectional regulatory loop between oocyte and cumulus cells. Our studies suggest that maintaining high cAMP levels in the oocyte during the first half of IVM sustained improved oocyte maturation, resulting in an enhanced response after IVF and cumulus matrix disassembly. Recent research indicated that the addition of OSFs during IVM enhanced the developmental competence of small follicle-derived oocytes, which was stimulated by epidermal growth factor (EGF) via developing EGF-receptor signaling. PMID:27349308

  4. Porcine oocyte maturation in vitro: role of cAMP and oocyte-secreted factors - A practical approach.

    Science.gov (United States)

    Appeltant, Ruth; Somfai, Tamás; Maes, Dominiek; VAN Soom, Ann; Kikuchi, Kazuhiro

    2016-10-18

    Polyspermy or the penetration of more than one sperm cell remains a problem during porcine in vitro fertilization (IVF). After in vitro culture of porcine zygotes, only a low percentage of blastocysts develop and their quality is inferior to that of in vivo derived blastocysts. It is unknown whether the cytoplasmic maturation of the oocyte is sufficiently sustained in current in vitro maturation (IVM) procedures. The complex interplay between oocyte and cumulus cells during IVM is a key factor in this process. By focusing on this bidirectional communication, it is possible to control the coordination of cumulus expansion, and nuclear and cytoplasmic maturation during IVM to some extent. Therefore, this review focuses on the regulatory mechanisms between oocytes and cumulus cells to further the development of new in vitro embryo production (IVP) procedures, resulting in less polyspermy and improved oocyte developmental potential. Specifically, we focused on the involvement of cAMP in maturation regulation and function of oocyte-secreted factors (OSFs) in the bidirectional regulatory loop between oocyte and cumulus cells. Our studies suggest that maintaining high cAMP levels in the oocyte during the first half of IVM sustained improved oocyte maturation, resulting in an enhanced response after IVF and cumulus matrix disassembly. Recent research indicated that the addition of OSFs during IVM enhanced the developmental competence of small follicle-derived oocytes, which was stimulated by epidermal growth factor (EGF) via developing EGF-receptor signaling.

  5. A Workflow to Investigate Exposure and Pharmacokinetic Influences on High-Throughput in Vitro Chemical Screening Based on Adverse Outcome Pathways.

    Science.gov (United States)

    Phillips, Martin B; Leonard, Jeremy A; Grulke, Christopher M; Chang, Daniel T; Edwards, Stephen W; Brooks, Raina; Goldsmith, Michael-Rock; El-Masri, Hisham; Tan, Yu-Mei

    2016-01-01

    Adverse outcome pathways (AOPs) link adverse effects in individuals or populations to a molecular initiating event (MIE) that can be quantified using in vitro methods. Practical application of AOPs in chemical-specific risk assessment requires incorporation of knowledge on exposure, along with absorption, distribution, metabolism, and excretion (ADME) properties of chemicals. We developed a conceptual workflow to examine exposure and ADME properties in relation to an MIE. The utility of this workflow was evaluated using a previously established AOP, acetylcholinesterase (AChE) inhibition. Thirty chemicals found to inhibit human AChE in the ToxCast™ assay were examined with respect to their exposure, absorption potential, and ability to cross the blood-brain barrier (BBB). Structures of active chemicals were compared against structures of 1,029 inactive chemicals to detect possible parent compounds that might have active metabolites. Application of the workflow screened 10 "low-priority" chemicals of 30 active chemicals. Fifty-two of the 1,029 inactive chemicals exhibited a similarity threshold of ≥ 75% with their nearest active neighbors. Of these 52 compounds, 30 were excluded due to poor absorption or distribution. The remaining 22 compounds may inhibit AChE in vivo either directly or as a result of metabolic activation. The incorporation of exposure and ADME properties into the conceptual workflow eliminated 10 "low-priority" chemicals that may otherwise have undergone additional, resource-consuming analyses. Our workflow also increased confidence in interpretation of in vitro results by identifying possible "false negatives." Phillips MB, Leonard JA, Grulke CM, Chang DT, Edwards SW, Brooks R, Goldsmith MR, El-Masri H, Tan YM. 2016. A workflow to investigate exposure and pharmacokinetic influences on high-throughput in vitro chemical screening based on adverse outcome pathways. Environ Health Perspect 124:53-60; http://dx.doi.org/10.1289/ehp.1409450.

  6. Correlating coating characteristics with the performance of drug-coated balloons--a comparative in vitro investigation of own established hydrogel- and ionic liquid-based coating matrices.

    Directory of Open Access Journals (Sweden)

    Sebastian Kaule

    Full Text Available Drug-coated balloons (DCB, which have emerged as a therapeutic alternative to drug-eluting stents in percutaneous cardiovascular intervention, are well described with regard to clinical efficacy and safety within a number of clinical studies. In vitro studies elucidating the correlation between coating additive and DCB performance are however rare but considered important for the understanding of DCB requirements and the improvement of established DCB. In this regard, we examined three different DCB-systems, which were developed in former studies based on the ionic liquid cetylpyridinium salicylate, the body-own hydrogel hyaluronic acid and the pharmaceutically well-established hydrogel polyvinylpyrrolidone, considering coating morphology, coating thickness, drug-loss, drug-transfer to the vessel wall, residual drug-concentration on the balloon surface and entire drug-load during simulated use in an in vitro vessel model. Moreover, we investigated particle release of the different DCB during simulated use and determined the influence of the three coatings on the mechanical behavior of the balloon catheter. We could show that coating characteristics can be indeed correlated with the performance of DCB. For instance, paclitaxel incorporation in the matrix can reduce the drug wash-off and benefit a high drug transfer. Additionally, a thin coating with a smooth surface and high but delayed solubility can reduce drug wash-off and decrease particle burden. As a result, we suggest that it is very important to characterize DCB in terms of mentioned properties in vitro in addition to their clinical efficacy in order to better understand their function and provide more data for the clinicians to improve the tool of DCB in coronary angioplasty.

  7. Correlating Coating Characteristics with the Performance of Drug-Coated Balloons – A Comparative In Vitro Investigation of Own Established Hydrogel- and Ionic Liquid-Based Coating Matrices

    Science.gov (United States)

    Kaule, Sebastian; Minrath, Ingo; Stein, Florian; Kragl, Udo; Schmidt, Wolfram; Schmitz, Klaus-Peter; Sternberg, Katrin; Petersen, Svea

    2015-01-01

    Drug-coated balloons (DCB), which have emerged as a therapeutic alternative to drug-eluting stents in percutaneous cardiovascular intervention, are well described with regard to clinical efficacy and safety within a number of clinical studies. In vitro studies elucidating the correlation between coating additive and DCB performance are however rare but considered important for the understanding of DCB requirements and the improvement of established DCB. In this regard, we examined three different DCB-systems, which were developed in former studies based on the ionic liquid cetylpyridinium salicylate, the body-own hydrogel hyaluronic acid and the pharmaceutically well-established hydrogel polyvinylpyrrolidone, considering coating morphology, coating thickness, drug-loss, drug-transfer to the vessel wall, residual drug-concentration on the balloon surface and entire drug-load during simulated use in an in vitro vessel model. Moreover, we investigated particle release of the different DCB during simulated use and determined the influence of the three coatings on the mechanical behavior of the balloon catheter. We could show that coating characteristics can be indeed correlated with the performance of DCB. For instance, paclitaxel incorporation in the matrix can reduce the drug wash-off and benefit a high drug transfer. Additionally, a thin coating with a smooth surface and high but delayed solubility can reduce drug wash-off and decrease particle burden. As a result, we suggest that it is very important to characterize DCB in terms of mentioned properties in vitro in addition to their clinical efficacy in order to better understand their function and provide more data for the clinicians to improve the tool of DCB in coronary angioplasty. PMID:25734818

  8. Correlating coating characteristics with the performance of drug-coated balloons--a comparative in vitro investigation of own established hydrogel- and ionic liquid-based coating matrices.

    Science.gov (United States)

    Kaule, Sebastian; Minrath, Ingo; Stein, Florian; Kragl, Udo; Schmidt, Wolfram; Schmitz, Klaus-Peter; Sternberg, Katrin; Petersen, Svea

    2015-01-01

    Drug-coated balloons (DCB), which have emerged as a therapeutic alternative to drug-eluting stents in percutaneous cardiovascular intervention, are well described with regard to clinical efficacy and safety within a number of clinical studies. In vitro studies elucidating the correlation between coating additive and DCB performance are however rare but considered important for the understanding of DCB requirements and the improvement of established DCB. In this regard, we examined three different DCB-systems, which were developed in former studies based on the ionic liquid cetylpyridinium salicylate, the body-own hydrogel hyaluronic acid and the pharmaceutically well-established hydrogel polyvinylpyrrolidone, considering coating morphology, coating thickness, drug-loss, drug-transfer to the vessel wall, residual drug-concentration on the balloon surface and entire drug-load during simulated use in an in vitro vessel model. Moreover, we investigated particle release of the different DCB during simulated use and determined the influence of the three coatings on the mechanical behavior of the balloon catheter. We could show that coating characteristics can be indeed correlated with the performance of DCB. For instance, paclitaxel incorporation in the matrix can reduce the drug wash-off and benefit a high drug transfer. Additionally, a thin coating with a smooth surface and high but delayed solubility can reduce drug wash-off and decrease particle burden. As a result, we suggest that it is very important to characterize DCB in terms of mentioned properties in vitro in addition to their clinical efficacy in order to better understand their function and provide more data for the clinicians to improve the tool of DCB in coronary angioplasty.

  9. The investigational Aurora kinase A inhibitor MLN8237 induces defects in cell viability and cell-cycle progression in malignant bladder cancer cells in vitro and in vivo.

    Science.gov (United States)

    Zhou, Ning; Singh, Kamini; Mir, Maria C; Parker, Yvonne; Lindner, Daniel; Dreicer, Robert; Ecsedy, Jeffrey A; Zhang, Zhongfa; Teh, Bin T; Almasan, Alexandru; Hansel, Donna E

    2013-04-01

    Despite more than 70,000 new cases of bladder cancer in the United States annually, patients with advanced disease have a poor prognosis due to limited treatment modalities. We evaluated Aurora kinase A, identified as an upregulated candidate molecule in bladder cancer, as a potential therapeutic target. Gene expression in human bladder cancer samples was evaluated using RNA microarray and quantitative reverse transcriptase PCR. Effects of the Aurora kinase A inhibitor MLN8237 (Millennium) on cell dynamics in malignant T24 and UM-UC-3 and papilloma-derived RT4 bladder cells were evaluated in vitro and in vivo in a mouse xenograft model. A set of 13 genes involved in the mitotic spindle checkpoint, including Aurora kinases A and B, were upregulated in human urothelial carcinoma compared with normal urothelium. The Aurora kinase A inhibitor MLN8237 induced cell-cycle arrest, aneuploidy, mitotic spindle failure, and apoptosis in the human bladder cancer cell lines T24 and UM-UC-3. MLN8237 also arrested tumor growth when administered orally over 4 weeks in a mouse bladder cancer xenograft model. Finally, in vitro sequential administration of MLN8237 with either paclitaxel or gemcitabine resulted in synergistic cytotoxic effects in T24 cells. Mitotic spindle checkpoint dysfunction is a common characteristic of human urothelial carcinoma and can be exploited with pharmacologic Aurora A inhibition. Given our demonstration of the ability of the Aurora A inhibitor MLN8237 to inhibit growth of bladder cancer in vitro and in vivo, we conclude that Aurora kinase inhibitors warrant further therapeutic investigation in bladder cancer. ©2013 AACR.

  10. The investigational Aurora kinase A inhibitor MLN8237 induces defects in cell viability and cell cycle progression in malignant bladder cancer cells in vitro and in vivo

    Science.gov (United States)

    Zhou, Ning; Singh, Kamini; Mir, Maria C.; Parker, Yvonne; Lindner, Daniel; Dreicer, Robert; Ecsedy, Jeffrey A; Zhang, Zhongfa; Teh, Bin T.; Almasan, Alexandru; Hansel, Donna E.

    2013-01-01

    PURPOSE Despite over 70,000 new cases of bladder cancer in the United States annually, patients with advanced disease have a poor prognosis due to limited treatment modalities. We evaluated Aurora kinase A, identified as an upregulated candidate molecule in bladder cancer, as a potential therapeutic target. EXPERIMENTAL DESIGN Gene expression in human bladder cancer samples was evaluated using RNA microarray and quantitative reverse-transcriptase PCR. Effects of the Aurora kinase A inhibitor MLN8237 (Millennium) on cell dynamics in malignant T24 and UM-UC-3 and papilloma-derived RT4 bladder cells were evaluated in vitro and in vivo in a mouse xenograft model. RESULTS A set of 13 genes involved in the mitotic spindle checkpoint, including Aurora kinases A and B, were upregulated in human urothelial carcinoma compared to normal urothelium. The Aurora kinase A inhibitor MLN8237 induced cell cycle arrest, aneuploidy, mitotic spindle failure, and apoptosis in the human bladder cancer cell lines T24 and UM-UC-3. MLN8237 also arrested tumor growth when administered orally over 4 weeks in a mouse bladder cancer xenograft model. Finally, in vitro sequential administration of MLN8237 with either paclitaxel or gemcitabine resulted in synergistic cytotoxic effects in T24 cells. CONCLUSIONS Mitotic spindle checkpoint dysfunction is a common characteristic of human urothelial carcinoma, and can be exploited with pharmacologic Aurora A inhibition. Given our demonstration of the ability of the Aurora A inhibitor MLN8237 to inhibit growth of bladder cancer in vitro and in vivo, we conclude that Aurora kinase inhibitors warrant further therapeutic investigation in bladder cancer. PMID:23403633

  11. Experimental approach to investigate the constrained recovery behavior of coiled monofilament polymer fibers

    Science.gov (United States)

    Mendes, S. S.; Nunes, L. C. S.

    2017-11-01

    The aim of this work is to propose a new approach for investigating the thermo-mechanical behavior of coiled oriented polymer fibers with fixed ends and promote an understanding of the actuation response of coiled polymers in constrained recovery applications. In the proposed experimental methodology, a coiled fiber was pre-stretched by 50% and the distance between its ends remained constant, then it was subjected to a heating-cooling cycle ranging from 30 °C to 120 °C and the induced restoring force was measured. Based on these measurements, axial deformation and shear strain were obtained from full-field displacements extracted by the digital image correlation method from images of the coiled fiber. Three coiled fibers with different initial pitch angles were manufactured, and samples with lengths of 15 mm and 20 mm were tested. Bias angles and coil radius were also estimated using the experimental data associated with the helical spring theory. Results show that significant shape changes can be noticed above the glass transition temperature (47 °C), and these changes induce variation in the resultant forces. The effects of thermal softening and thermal contraction for a modest negative thermal expansion coefficient became evident at temperatures ranging from ∼47 °C to ∼90 °C, while the response of a coiled homochiral polymer fiber was achieved at temperatures close to 90 °C. During the cooling process, saturated states of the axial deformation and shear strain of the coiled fibers were observed at temperatures between 120 °C and 100 °C.

  12. Rumination and implicit avoidance following bereavement: an approach avoidance task investigation.

    Science.gov (United States)

    Eisma, Maarten C; Rinck, Mike; Stroebe, Margaret S; Schut, Henk A W; Boelen, Paul A; Stroebe, Wolfgang; van den Bout, Jan

    2015-06-01

    Rumination, a risk factor in adjustment to bereavement, has often been considered a confrontation process. However, building on research on worry in generalized anxiety disorder (GAD) and rumination in post-traumatic stress disorder (PTSD), researchers recently developed the Rumination as Avoidance Hypothesis (RAH), which states that rumination after bereavement serves to avoid the reality of the loss. In the present study, RAH was tested by investigating if rumination is associated with implicit loss avoidance. An Approach Avoidance Task (AAT) was used to assess automatic behavior tendencies. Using a joystick, 71 persons who recently lost a first-degree relative (90.1% women), pulled stimuli toward themselves or pushed them away from themselves. Stimuli represented the loss (picture deceased + loss word), were loss-related but ambiguous (picture deceased + neutral word; picture stranger + loss word), or were non-loss-related (picture stranger + neutral word; puzzle picture + X's). Participants who ruminated more were relatively faster in pushing loss stimuli away from themselves and slower in pulling loss stimuli towards themselves, implying more rumination was associated with stronger implicit loss avoidance. Effects were maintained after controlling for depressive or post-traumatic stress symptom levels, but not when controlling for prolonged grief symptom levels. Conjugally bereaved women were overrepresented in the sample, which limits generalizability of results. The study was correlational, precluding causal inferences. In line with RAH, rumination was positively associated with loss avoidance. This may indicate that the application of exposure-based techniques can reduce rumination and loss-related psychopathology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Structural Investigation for Optimization of Anthranilic Acid Derivatives as Partial FXR Agonists by in Silico Approaches

    Directory of Open Access Journals (Sweden)

    Meimei Chen

    2016-04-01

    Full Text Available In this paper, a three level in silico approach was applied to investigate some important structural and physicochemical aspects of a series of anthranilic acid derivatives (AAD newly identified as potent partial farnesoid X receptor (FXR agonists. Initially, both two and three-dimensional quantitative structure activity relationship (2D- and 3D-QSAR studies were performed based on such AAD by a stepwise technology combined with multiple linear regression and comparative molecular field analysis. The obtained 2D-QSAR model gave a high predictive ability (R2train = 0.935, R2test = 0.902, Q2LOO = 0.899. It also uncovered that number of rotatable single bonds (b_rotN, relative negative partial charges (RPC−, oprea's lead-like (opr_leadlike, subdivided van der Waal’s surface area (SlogP_VSA2 and accessible surface area (ASA were important features in defining activity. Additionally, the derived3D-QSAR model presented a higher predictive ability (R2train = 0.944, R2test = 0.892, Q2LOO = 0.802. Meanwhile, the derived contour maps from the 3D-QSAR model revealed the significant structural features (steric and electronic effects required for improving FXR agonist activity. Finally, nine newly designed AAD with higher predicted EC50 values than the known template compound were docked into the FXR active site. The excellent molecular binding patterns of these molecules also suggested that they can be robust and potent partial FXR agonists in agreement with the QSAR results. Overall, these derived models may help to identify and design novel AAD with better FXR agonist activity.

  14. Investigating performance variability of processing, exploitation, and dissemination using a socio-technical systems analysis approach

    Science.gov (United States)

    Danczyk, Jennifer; Wollocko, Arthur; Farry, Michael; Voshell, Martin

    2016-05-01

    Data collection processes supporting Intelligence, Surveillance, and Reconnaissance (ISR) missions have recently undergone a technological transition accomplished by investment in sensor platforms. Various agencies have made these investments to increase the resolution, duration, and quality of data collection, to provide more relevant and recent data to warfighters. However, while sensor improvements have increased the volume of high-resolution data, they often fail to improve situational awareness and actionable intelligence for the warfighter because it lacks efficient Processing, Exploitation, and Dissemination and filtering methods for mission-relevant information needs. The volume of collected ISR data often overwhelms manual and automated processes in modern analysis enterprises, resulting in underexploited data, insufficient, or lack of answers to information requests. The outcome is a significant breakdown in the analytical workflow. To cope with this data overload, many intelligence organizations have sought to re-organize their general staffing requirements and workflows to enhance team communication and coordination, with hopes of exploiting as much high-value data as possible and understanding the value of actionable intelligence well before its relevance has passed. Through this effort we have taken a scholarly approach to this problem by studying the evolution of Processing, Exploitation, and Dissemination, with a specific focus on the Army's most recent evolutions using the Functional Resonance Analysis Method. This method investigates socio-technical processes by analyzing their intended functions and aspects to determine performance variabilities. Gaps are identified and recommendations about force structure and future R and D priorities to increase the throughput of the intelligence enterprise are discussed.

  15. Investigation of a common gene expression signature in gastrointestinal cancers using systems biology approaches.

    Science.gov (United States)

    Baghaei, Kaveh; Hosseinkhan, Nazanin; Asadzadeh Aghdaei, Hamid; Zali, M R

    2017-10-24

    According to GLOBOCAN 2012, the incidence and the mortality rate of colorectal, stomach and liver cancers are the highest among the total gastrointestinal (GI) cancers. Here we aimed to find the common genes and pathways that are simultaneously deregulated in these three malignancies using systems biology approaches. Here we conducted a differential expression analysis on high-quality gene expression datasets of gastric cancer (GC), colorectal cancer (CRC) and hepatocellular carcinoma (HCC). To address the inter gene correlations that were neglected in differential expression studies, we also applied differential co-expression analysis on the understudied datasets. The common significant differentially expressed genes (DEGs) among the three cancers were used for further regulatory and PPI network construction. In parallel the regulatory roles of miRNAs and lncRNAs in the common DEGs were investigated. 23 common DEGs were detected between GC, CRC and HCC. Two cases of potential feed forward loops were identified in the constructed TF-target regulatory network, indicating the probable cross-talk between biological pathways. The result of a vulnerability test on the common PPI network resulted in the finding of three candidates, the simultaneous targeting of which will disintegrate the main parts of the network. The results of the differential co-expression study led to the identification of respectively 7 and 1 common differentially co-expressed pairs of genes between GC and CRC and between CRC and HCC. The results of the differential expression study introduced new common players in CRC, GC and HCC and provided better insights into the molecular characteristics of these GI malignancies. Moreover, we concluded that differential co-expression studies are an essential complement for differential expression studies that just take single differentially expressed genes into account.

  16. Investigation of Liver Injury of Polygonum multiflorum Thunb. in Rats by Metabolomics and Traditional Approaches

    Science.gov (United States)

    Li, Yun-Xia; Gong, Xiao-Hong; Liu, Mei-Chen; Peng, Cheng; Li, Peng; Wang, Yi-Tao

    2017-01-01

    Liver injury induced by Polygonum multiflorum Thunb. (PM) have been reported since 2006, which aroused widespread concern. However, the toxicity mechanism of PM liver injury remained unclear. In this study, the mechanism of liver injury induced by different doses of PM after long-term administration was investigated in rats by metabolomics and traditional approaches. Rats were randomly divided into control group and PM groups. PM groups were oral administered PM of low (10 g/kg), medium (20 g/kg), high (40 g/kg) dose, while control group was administered distilled water. After 28 days of continuous administration, the serum biochemical indexes in the control and three PM groups were measured and the liver histopathology were analyzed. Also, UPLC-Q-TOF-MS with untargeted metabolomics was performed to identify the possible metabolites and pathway of liver injury caused by PM. Compared with the control group, the serum levels of ALT, AST, ALP, TG, and TBA in middle and high dose PM groups were significantly increased. And the serum contents of T-Bil, D-Bil, TC, TP were significantly decreased. However, there was no significant difference between the low dose group of PM and the control group except serum AST, TG, T-Bil, and D-Bil. Nine biomarkers were identified based on biomarkers analysis. And the pathway analysis indicated that fat metabolism, amino acid metabolism and bile acid metabolism were involved in PM liver injury. Based on the biomarker pathway analysis, PM changed the lipid metabolism, amino acid metabolism and bile acid metabolism and excretion in a dose-dependent manner which was related to the mechanism of liver injury. PMID:29163173

  17. Investigation of Liver Injury of Polygonum multiflorum Thunb. in Rats by Metabolomics and Traditional Approaches

    Directory of Open Access Journals (Sweden)

    Yun-Xia Li

    2017-11-01

    Full Text Available Liver injury induced by Polygonum multiflorum Thunb. (PM have been reported since 2006, which aroused widespread concern. However, the toxicity mechanism of PM liver injury remained unclear. In this study, the mechanism of liver injury induced by different doses of PM after long-term administration was investigated in rats by metabolomics and traditional approaches. Rats were randomly divided into control group and PM groups. PM groups were oral administered PM of low (10 g/kg, medium (20 g/kg, high (40 g/kg dose, while control group was administered distilled water. After 28 days of continuous administration, the serum biochemical indexes in the control and three PM groups were measured and the liver histopathology were analyzed. Also, UPLC-Q-TOF-MS with untargeted metabolomics was performed to identify the possible metabolites and pathway of liver injury caused by PM. Compared with the control group, the serum levels of ALT, AST, ALP, TG, and TBA in middle and high dose PM groups were significantly increased. And the serum contents of T-Bil, D-Bil, TC, TP were significantly decreased. However, there was no significant difference between the low dose group of PM and the control group except serum AST, TG, T-Bil, and D-Bil. Nine biomarkers were identified based on biomarkers analysis. And the pathway analysis indicated that fat metabolism, amino acid metabolism and bile acid metabolism were involved in PM liver injury. Based on the biomarker pathway analysis, PM changed the lipid metabolism, amino acid metabolism and bile acid metabolism and excretion in a dose-dependent manner which was related to the mechanism of liver injury.

  18. BIOTECHNOLOGICAL APPROACHES FOR CONSERVATION OF THE ENDANGERED SPECIES Crambe koktebelica (JUNGE N. BUSCH AND EFFECT OF ASEPTIC IN VITRO CULTIVATION ON ITS BIOCHEMICAL PROPERTIES

    Directory of Open Access Journals (Sweden)

    Pushkarova

    2016-08-01

    Full Text Available The aim of the study was to establish efficient protocols of seed surface sterilization with further multiplication in vitro for threatened species Crambe koktebelica (Junge N. Busch and to show the effect of biotechnological approach (in vitro cultivation of biodiversity conservation on plants biochemical properties. Seed surface sterilization was carried out according to the original method with further microclonal multiplication of aseptic sprouts from lateral buds on the Murashige and Skoog (MS medium supplemented with different concentrations of growth regulators. Fatty acid content was determined using Gas chromatography-mass spectrophotometry of fatty acid ethers. Antioxidant activity was determined using 2.2-diphenyl-1-picrylhydrazyl radical scavenging method. Total soluble protein content was measured using Bradford method and polyfructan content determination was based upon ketosugars ability to color in the acidic environment with resorcinol. Plants that were grown under in vitro and in vivo conditions and seeds were used in this research. Efficient protocol of surface sterilization that resulted in 45% of aseptic seed material 50% of which has sprouted was elaborated for C. koktebelica as well as fast microclonal multiplication methods that provided with up to 5.25 ± 0.50 new formed plantlets from 1 lateral bud (on the MS medium that contained 1 mg/L of 6-benzylaminopurine. It was also shown that aseptic cultivation benefits to saturated fatty acid accumulation and increases protein content but on the other hand it reduces unsaturated fatty acid amount and polyfructan content as well as antioxidant activity of plant material. Obtained data confirms the prospect of biotechnology approach to biodiversity conservation and suggest the necessity of father in vitro cultivation effect on biochemical composition of plant study.

  19. An impedance-based approach using human iPSC-derived cardiomyocytes significantly improves in vitro prediction of in vivo cardiotox liabilities.

    Science.gov (United States)

    Koci, Bryan; Luerman, Gregory; Duenbostell, Anika; Kettenhofen, Ralf; Bohlen, Heribert; Coyle, Luke; Knight, Brian; Ku, Warren; Volberg, Walter; Woska, Joseph R; Brown, Martha P

    2017-08-15

    Current in vitro approaches to cardiac safety testing typically focus on mechanistic ion channel testing to predict in vivo proarrhythmic potential. Outside of the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative, structural and functional cardiotoxicity related to chronic dosing effects are of great concern as these effects can impact compound attrition. Development and implementation of an in vitro cardiotoxicity screening platform that effectively identifies these liabilities early in the discovery process should reduce costly attrition and decrease preclinical development time. Impedence platforms have the potential to accurately identify structural and functional cardiotoxicity and have sufficient throughput to be included in a multi-parametric optimization approach. Human induced pluripotent stem cell cardiomyocytes (hIPSC-CMs) have demonstrated utility in cardiac safety and toxicity screening. The work described here leverages these advantages to assess the predictive value of data generated by two impedance platforms. The response of hIPSC-CMs to compounds with known or predicted cardiac functional or structural toxicity was determined. The compounds elicited cardiac activities and/or effects on "macro" impedance often associated with overt structural or cellular toxicity, detachment, or hypertrophy. These assays correctly predicted in vivo cardiotox findings for 81% of the compounds tested and did not identify false positives. In addition, internal or literature Cmax values from in vivo studies correlated within 4 fold of the in vitro observations. The work presented here demonstrates the predictive power of impedance platforms with hIPSC-CMs and provides a means toward accelerating lead candidate selection by assessing preclinical cardiac safety earlier in the drug discovery process. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Quantitative Comparison of Three Endoscopic Approaches to the Parasellar Region: Laboratory Investigation.

    Science.gov (United States)

    Belykh, Evgenii; Yağmurlu, Kaan; Hong, Yuan; Mooney, Michael A; Bozkurt, Baran; Byvaltsev, Vadim A; Nakaji, Peter; Preul, Mark C

    2017-12-01

    Endoscopic endonasal transsphenoidal and contralateral sublabial transmaxillary approaches are used for approaching parasellar lesions. The aim of this anatomical study was to compare endoscopic endonasal uninostril and binostril (contralateral) and contralateral sublabial transmaxillary approaches via a quantitative analysis of exposure limits and instrument working avenues. Six formalin-fixed silicone-injected adult cadaveric heads (12 sides) were studied. The surgical working area, depth of the surgical corridor, angle of attack, and surgical freedom were measured and compared for the 3 approaches. The endoscopic binostril endonasal approach to the parasellar area provided greater surgical freedom in the opticocarotid recess (OCR) and superior orbital fissure (SOF) compared with that of the uninostril endonasal approach (OCR, P trajectory to the contralateral parasellar area (P < 0.01). An anatomical comparison of the 3 endoscopic approaches to the parasellar area showed that the binostril approach provides greater exposure and freedom for instrument manipulation. The contralateral transmaxillary route provided a more lateral view, increasing exposure on average by 48%, with shorter surgical depth; however, surgical freedom was inferior to that of the binostril approach. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. A functional genomics approach using radiation-induced changes in gene expression to study low dose radiation effects in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Fornace, Jr, A J

    2007-03-03

    Abstract for final report for project entitled A functional genomics approach using radiation-induced changes in gene expression to study low dose radiation effects in vitro and in vivo which has been supported by the DOE Low Dose Radiation Research Program for approximately 7 years. This project has encompassed two sequential awards, ER62683 and then ER63308, in the Gene Response Section in the Center for Cancer Research at the National Cancer Institute. The project was temporarily suspended during the relocation of the Principal Investigators laboratory to the Dept. of Genetics and Complex Diseases at Harvard School of Public Health at the end of 2004. Remaining support for the final year was transferred to this new site later in 2005 and was assigned the DOE Award Number ER64065. The major aims of this project have been 1) to characterize changes in gene expression in response to low-dose radiation responses; this includes responses in human cells lines, peripheral blood lymphocytes (PBL), and in vivo after human or murine exposures, as well as the effect of dose-rate on gene responses; 2) to characterize changes in gene expression that may be involved in bystander effects, such as may be mediated by cytokines and other intercellular signaling proteins; and 3) to characterize responses in transgenic mouse models with relevance to genomic stability. A variety of approaches have been used to study transcriptional events including microarray hybridization, quantitative single-probe hybridization which was developed in this laboratory, quantitative RT-PCR, and promoter microarray analysis using genomic regulatory motifs. Considering the frequent responsiveness of genes encoding cytokines and related signaling proteins that can affect cellular metabolism, initial efforts were initiated to study radiation responses at the metabolomic level and to correlate with radiation-responsive gene expression. Productivity includes twenty-four published and in press manuscripts

  2. Investigation of in vitro and in vivo antioxidant activities of flavonoids rich extract from the berries of Rhodomyrtus tomentosa(Ait.) Hassk.

    Science.gov (United States)

    Wu, Pingping; Ma, Guangzhi; Li, Nianghui; Deng, Qian; Yin, Yanyan; Huang, Ruqiang

    2015-04-15

    This study investigated the in vitro and in vivo antioxidant activities of the flavonoids rich extract from Rhodomyrtus tomentosa Hassk (R. tomentosa) berries. The in vitro antioxidant assay demonstrated that the flavonoids rich extract (62.09% rutin equivalent) extracted by ethanol and purified by AB-8 macroporous resin was strong in reducing power, superoxide radical, hydroxyl radical and DPPH radical scavenging activity, as well as inhibiting lipid peroxidation. In the in vivo assays, the flavonoids rich extract significantly enhanced the activities of antioxidant enzymes in serums of mice after they were administered with the extract. The results suggested that the flavonoids rich extract from R. tomentosa fruits possesses potent antioxidant properties. In addition, the chemical compositions of flavonoids rich extract were identified by UPLC-TOF-MS/MS. Six flavonoids were tentatively identified as myricetin, quercetin, dihydromyricetin, kaempferol, quercetin 7,4'-diglucoside and vitexin. Therefore, R. tomentosa berries could be used as a new source of antioxidant ingredient. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Nanoparticle Fullerene (C60) demonstrated stable binding with antibacterial potential towards probable targets of drug resistant Salmonella typhi - a computational perspective and in vitro investigation.

    Science.gov (United States)

    Skariyachan, Sinosh; Parveen, Asma; Garka, Shruti

    2017-12-01

    Salmonella typhi, a Gram negative bacterium, has become multidrug resistant (MDR) to wide classes of antibacterials which necessitate an alarming precaution. This study focuses on the binding potential and therapeutic insight of Nano-Fullerene C60 towards virulent targets of Salmonella typhi by computational prediction and preliminary in vitro assays. The clinical isolates of Salmonella typhi were collected and antibiotic susceptibility profiles were assessed. The drug targets of pathogen were selected by rigorous literature survey and gene network analysis by various metabolic network resources. Based on this study, 20 targets were screened and the 3D structures of few drug targets were retrieved from PDB and others were computationally predicted. The structures of nanoleads such as Fullerene C60, ZnO and CuO were retrieved from drug databases. The binding potential of these nanoleads towards all selected targets were predicted by molecular docking. The best docked conformations were screened and concept was investigated by preliminary bioassays. This study revealed that most of the isolates of Salmonella typhi were found to be MDR (p C60 showed better binding affinity towards the drug targets when compared to ZnO and CuO. The preliminary in vitro assays suggested that 100 μg/L Fullerene C60 posses significant inhibitory activities and absence of drug resistance to this nanoparticle. This study suggests that Fullerene C60 can be scaled up as probable lead molecules against the major drug targets of MDR Salmonella typhi.

  4. Identification and investigation on antagonistic effect of Trichoderma spp. on tea seedlings white foot and root rot (Sclerotium rolfsii Sacc.) in vitro condition.

    Science.gov (United States)

    Shaigan, S; Seraji, A; Moghaddam, S A M

    2008-10-01

    The aim of this two years research work is to recognize bio control agents and its antagonistic efficiency on Sclerotium rolfsii in vitro condition. Sclerotium rolfsii Sacc. is a causal agent of white foot and root rot disease in many plants. This fungi causing damage in tea nurseries of tea cultivated countries especially in Iran. Due to importance this disease in tea nurseries of Iran and impossibility using of chemical control against this damaging agent In this research, five species of Trichoderma viride, Trichoderma harzianum, Trichoderma hamatum, Trichoderma longibrachiatum and Trichoderma parceramosum as antagonist fungi against Sclerotium rolfsii were collected and identified from tea nurseries and seedlings in Iran. Then efficiency of these bio control agent against Sclerotium rolfsii were investigated in vitro condition. Results showed that these species with different mechanism such as lysis of sclerotia, inhibited mycelial growth of Sclerotium rolfsii with volatile metabolites producing and parasitized the hyphal trends of disease agent were showed its antagonistic effects against causal agent of white foot and root rot in tea seedling. Trichoderma viride, Trichoderma harzianum and Trichoderma hamatum after 30 days destructed and lysis the sclerotia 98.5, 86.5 and 85%, respectively. Producing of volatile metabolites after 72 h reciprocal growth of Trichoderma viride, Trichoderma harzianum and Trichoderma longibrachiatum till 60.8, 54.8 and 54.4% prevented mycelium growth of Sclerotium rolfsii, respectively. All identified species caused parasitize (to hyphal contact method) pathogen fungal hypha that the first three species showed more parasite effect.

  5. In vitro investigation of orthopedic titanium-coated and brushite-coated surfaces using human osteoblasts in the presence of gentamycin.

    Science.gov (United States)

    Ince, Akif; Schütze, Norbert; Hendrich, Christian; Thull, Roger; Eulert, Jochen; Löhr, Jochen F

    2008-08-01

    Anti-infective coatings have been developed to protect the surfaces of cementless implants from bacterial colonization that is known to be a prerequisite for device-related infection. The aim of this study is to investigate the effect of brushite-coated arthroplasty surfaces on human osteoblasts and to evaluate the impact of concomitant exposure to gentamycin. We cultured human osteoblasts (hFOB 1.19) on brushite-coated and uncoated titanium alloy in the presence of gentamycin and analyzed cell function and vitality. Our results show that brushite-coated titanium alloy surfaces supported the function of osteoblasts and the expression of extracellular matrix even in the presence of highly dosed gentamycin. Brushite-coated titanium alloy surfaces supported osteogenic function, indicating that this coating could enhance implant osteointegration in vivo. Concomitant exposure to gentamycin slightly decreased osteoblastic activity in vitro, suggesting that there might also be negative effects in vivo. However, in vivo studies are necessary to validate these in vitro findings.

  6. Investigating the Process Approach to Writing Instruction in Urban Middle Schools

    Science.gov (United States)

    Patthey-Chavez, G. Genevieve; Matsumura, Lindsay Clare; Valdes, Rosa

    2004-01-01

    The process approach to writing instruction emphasizes a cycle of revision during which students draft, edit, revise, and redraft their work. In this approach, feedback from teachers or peers and the opportunity to revise written work based on this feedback are considered to be keys to students' development as writers, and the role of instruction…

  7. Investigating DIF and extensions using an LLTM approach and also an individual differences approach: an international testing context

    Directory of Open Access Journals (Sweden)

    YIYU XIE

    2008-09-01

    Full Text Available This study intends to investigate two ways to generalise differential item functioning (DIF by grouping of items that share a common feature, or an item property as in the Linear Logistic Test Model (LLTM. An item “facet” refers to this type of grouping, and DIF can be expressed in terms of more fundamental parameters that relate to the facet of items. Hence the differential facet functioning (DFF model, a particular version of the LLTM, helps to explain the DIF effects more substantively. Using the mathematics data from the Program for International Student Assessment (PISA 2003, this study shows that modeling the DFF effect through an interaction of the group-by-facet parameter rather than DIF effect on the individual item level can be handled easily with the NLMIXED procedure of SAS. We found that the results are more interpretable when the bias is interpreted on the facet level rather than the item level. Analogous to the multidimensional DIF model, one natural extension of the DFF model is to make the model multidimensional when DFF facets (i.e., LLTM facets are considered as dimensions. This extension, multidimensional DFF (MDFF, is also investigated. The MDFF model allows individual differences to be modeled on the dimension that exhibits a DFF effect. However, it is always recommended to check the individual DIF estimates and construct a substantive analysis first before conducting DFF and MDFF analysis.

  8. Inhibition of in vitro metabolism of testosterone in human, dog and horse liver microsomes to investigate species differences.

    Science.gov (United States)

    Zielinski, Jana; Mevissen, Meike

    2015-04-01

    Testosterone hydroxylation was investigated in human, canine and equine liver microsomes and in human and canine single CYPs. The contribution of the CYP families 1, 2 and 3 was studied using chemical inhibitors. Testosterone metabolites were analyzed by HPLC. The metabolites androstenedione, 6β- and 11β-hydroxytestosterone were found in microsomes of all species, but the pattern of metabolites varied within species. Androstenedione was more prominent in the animal species, and an increase over time was seen in equines. Testosterone hydroxylation was predominantly catalyzed by the CYP3A subfamily in all three species. While CYP2C9 did not metabolise testosterone, the canine ortholog CYP2C21 produced androstenedione. Quercetin significantly inhibited 6β- and 11β-hydroxytestosterone in all species investigated, suggesting that CYP2C8 is involved in testosterone metabolism, whereas sulfaphenazole significantly inhibited the formation of 6β- and 11β-hydroxytestosterone in human microsomes, at 60 min in equine microsomes, but not in canine microsomes. A contribution of CYP2B6 in testosterone metabolism was only found in human and equine microsomes. Inhibition of 17β-hydroxysteroid dehydrogenase 2 indicated its involvement in androstenedione formation in humans, increased androstenedione formation was found in equines and no involvement in canines. These findings provide improved understanding of differences in testosterone biotransformation in animal species. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Hall versus conventional stainless steel crown techniques: in vitro investigation of marginal fit and microleakage using three different luting agents.

    Science.gov (United States)

    Erdemci, Zeynep Yalçınkaya; Cehreli, S Burçak; Tirali, R Ebru

    2014-01-01

    This study's purpose was to investigate microleakage and marginal discrepancies in stainless steel crowns (SSCs) placed using conventional and Hall techniques and cemented with three different luting agents. Seventy-eight human primary maxillary second molars were randomly assigned to two groups (N=39), and SSCs were applied either with the Hall or conventional technique. These two groups were further subgrouped according to the material used for crown cementation (N=13 per group). Two specimens in each group were processed for scanning electron microscopy investigation. The extent of microleakage and marginal fit was quantified in millimeters on digitally photographed sections using image analysis software. The data were compared with a two-way independent and a two-way mixed analysis of variance (P=.05). The scores in the Hall group were significantly worse than those in the conventional technique group (PStainless steel crowns applied using the Hall technique displayed higher microleakage scores than those applied using the conventional technique, regardless of the cementation material. When the interaction of the material and technique was assessed, resin cement presented as the best choice for minimizing microleakage in both techniques.

  10. Investigation of Multi-Criteria Decision Consistency: A Triplex Approach to Optimal Oilfield Portfolio Investment Decisions

    Science.gov (United States)

    Qaradaghi, Mohammed

    Complexity of the capital intensive oil and gas portfolio investments is continuously growing. It is manifested in the constant increase in the type, number and degree of risks and uncertainties, which consequently lead to more challenging decision making problems. A typical complex decision making problem in petroleum exploration and production (E&P) is the selection and prioritization of oilfields/projects in a portfolio investment. Prioritizing oilfields maybe required for different purposes, including the achievement of a targeted production and allocation of limited available development resources. These resources cannot be distributed evenly nor can they be allocated based on the oilfield size or production capacity alone since various other factors need to be considered simultaneously. These factors may include subsurface complexity, size of reservoir, plateau production and needed infrastructure in addition to other issues of strategic concern, such as socio-economic, environmental and fiscal policies, particularly when the decision making involves governments or national oil companies. Therefore, it would be imperative to employ decision aiding tools that not only address these factors, but also incorporate the decision makers' preferences clearly and accurately. However, the tools commonly used in project portfolio selection and optimization, including intuitive approaches, vary in their focus and strength in addressing the different criteria involved in such decision problems. They are also disadvantaged by a number of drawbacks, which may include lacking the capacity to address multiple and interrelated criteria, uncertainty and risk, project relationship with regard to value contribution and optimum resource utilization, non-monetary attributes, decision maker's knowledge and expertise, in addition to varying levels of ease of use and other practical and theoretical drawbacks. These drawbacks have motivated researchers to investigate other tools and

  11. Risk factor investigation for cardiovascular health through WHO STEPS approach in Ardabil, Iran

    Directory of Open Access Journals (Sweden)

    Mashadi-Abdollahi H

    2011-07-01

    Full Text Available H Sadeghi-Bazargani1,2, H Jafarzadeh3, M Fallah4, S Hekmat3, J Bashiri3, GH Hosseingolizadeh3, MS Soltanmohammadzadeh3, A Mortezazadeh3, A Shaker3, M Danehzan3, A Zohouri3, O Khosravi3, R Nasimidoust3, N Malekpour3, E Kharazmi4, M Babaei3, M Nadirmohammadi3, H Mashhadi-Abdollahi51Neuroscience Research Center, 2Statistics and Epidemiology Department, Faculty of Health and Nutrition, Tabriz University of Medical Sciences, Tabriz, Iran; 3Public Health Department, Ardabil University of Medical Sciences, Ardabil, Iran; 4Tampere School of Public Health, University of Tampere, Tampere, Finland; 5National Public Health Management Center (NPMC, Tabriz University of Medical Sciences, Tabriz, IranObjectives: Reliable evidence is the keystone for any noncommunicable disease (NCD prevention plan to be initiated. In this study we carried out a risk factor investigation based on the WHO Stepwise approach to Surveillance (STEPS.Methods: The study was conducted out on 1000 adults between 15 and 64 years of age living in Ardabil province, north-west Iran during 2006, based on the WHO STEPS approach to surveillance of risk factors for NCD. At this stage only the first and second steps were carried out. Data were collected through standard questionnaires and methods analyzed using STATA version 8 statistical software package.Results: 29.0% of men and 2.6% of women were current daily tobacco smokers. The mean number of manufactured cigarettes smoked per day was 18.9 among current daily smokers. Smoking was most prevalent among men of low-income families and those of lower education. The mean body mass index (BMI was 26.6 kg/m2, and was significantly correlated with systolic blood pressure. 58.9% were overweight or obese; 18.0% had raised blood pressure and 3.7% had isolated systolic hypertension. The mean number of servings of fruit consumed per day was 1.1; 33.1% had low levels of activity. Combined risk factor analysis showed that 4.1% of participants were in the

  12. Integrated monitoring approach to investigate the contamination, mobilization and risks of sediments

    Science.gov (United States)

    Bölscher, Jens; Schulte, Achim; Terytze, Konstantin

    2017-04-01

    The use of surface water bodies for manufacturing purposes has been common not only in Germany since the beginning of industrialization, and this has led to a high accumulation of different chemical contaminants in the sediments of aquatic ecosystems. In particular, water bodies with very low flow conditions like the "Rummelsburger See", an anabranch of the Spree River located in the centre of Berlin, have been highly affected. Given that, it has become necessary to obtain improved knowledge concerning the current sediment dynamics, the rate of sedimentation and the current level of contamination and toxicity compared to earlier conditions. Against this background, a survey was set up, consisting of an integrated monitoring approach that focuses on hydraulics, sediment dynamics and contamination, including boundary conditions, such as weather and motor-boat activities to find information, which would help design appropriate treatment in the future. To detect the spatial distribution of pollutants in the sediment, over 200 sediment samples were collected via drill cores at 16 locations. The upper 15 cm of each drill core was systematically divided into 5 layers (each of 3 cm) for separate examination. The investigation of sediment deposition and remobilisation rates was accomplished by installing 18 sediment traps. The presence of selected heavy metals and organic pollutants in the sediments was determined for every sampling location and layer of the drill cores, as well as for all sediment traps. Changes in boundary conditions which influence the spatial and temporal distribution of deposition and resuspension were monitored by placing devices within the water body and taking different mobile measurements (3-D flow conditions, oxygen, turbidity, chlorophyll-a, temperature). The analysis of sediment and suspended matter included the determination of the total content of inorganic (Hg, Cd, Cr, Pb, Ni, Cu, Zn) and organic compounds (polycyclic aromatic hydrocarbons

  13. Mammalian cell display technology coupling with AID induced SHM in vitro: an ideal approach to the production of therapeutic antibodies.

    Science.gov (United States)

    Qin, Chang-Fei; Li, Guan-Cheng

    2014-12-01

    Traditional antibody production technology within non-mammalian cell expression systems has shown many unsatisfactory properties for the development of therapeutic antibodies. Nevertheless, mammalian cell display technology reaps the benefits of producing full-length all human antibodies. Together with the developed cytidine deaminase induced in vitro somatic hypermutation technology, mammalian cell display technology provides the opportunity to produce high affinity antibodies that might be ideal for therapeutic application. This review was concentrated on the development of the mammalian cell display technology as well as the activation-induced cytidine deaminase induced in vitro somatic hypermutation technology and their applications for the production of therapeutic antibodies. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Enhanced both in vitro and in vivo kinetics by SLNs induced transdermal system of furosemide: A novel approach.

    Science.gov (United States)

    Mannam, Revathi; Yallamalli, Indira Muzib

    2017-11-28

    Furosemide is a potent diuretic agent used to treat pulmonary arterial hypertension. Variable dosage regimen and poor pharmacokinetic parameters has led to the development of transdermal drug delivery system. A patent on suitability of multi-lamellar structures for excellent transdermal delivery (US 0367475A1) has encouraged us to formulate the solid lipid nanoparticles (SLNs) induced transdermal systems of furosemide to enhance the kinetic properties without incorporating any penetration enhancer and rate limiting polymers. SLNs were prepared by hot homogenization and ultra-sonication method; optimization was done basing on entrapment efficiency and particle size. Optimized SLNs were incorporated in to transdermal patches by solvent casting method. In-vitro and in-vivo studies were carried out for characterization of transdermal patches. SLNs of F9 (GMS: Span 60: Pluronic F 68 in 6:2.5:0.2) were optimized for incorporating in to transdermal system (entrapment efficiency 94.5±0.045%, particle size 69.6±1.48 nm and in-vitro release 94.38±1.02%). Transdermal patches were formulated using combinations of hydrophilic and hydrophobic polymers to study the diffusion kinetics. Formulation FS1 (HPMC 4 parts) was optimized for further studies (in-vitro release 98.11±1.21% with flux of 58.726±0.023 µg/cm2/h) and no significant difference from ex-vivo permeation studies was observed. Drug release followed mixed order diffusion kinetics and super case -II transport mechanism. In-vivo pharmacokinetic data of SLNs induced transdermal system suggested a 3.6 times increase in AUC and 5.4 times increase in MRT when compared with oral route. The SLNs induced transdermal patch was found to beneficial in enhancing kinetic properties both in-vitro and in-vivo. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  15. A facilitative versus directive approach in training clinical skills? Investigating students' clinical performance and perceptions.

    Science.gov (United States)

    Berghmans, Inneke; Druine, Nathalie; Dochy, Filip; Struyven, Katrien

    2012-08-01

    Over the years, many medical school curricula have started implementing diverse student-centred teaching and learning methodologies. Previous studies, however, have indicated that students prefer more traditional and directive methodologies instead, raising questions on which training approach should be advocated. This study contrasts the effects of a student-centred (i.e. facilitative) training approach on students' clinical skills learning with students' perceptions. More specifically, a quasi-experimental study was set up in which students experienced either a directive or facilitative training approach. Data were collected by means of an OSCE on the one hand, and a questionnaire on students' perceptions of the training sessions, and two open-ended questions about students' likes and dislikes on the other hand. While no general differences were found in terms of clinical knowledge and understanding, and actual clinical performance, an interaction between students' course-specific prior knowledge and the training approach was found. Especially students with low levels of knowledge benefited more from the facilitative training approach in terms of clinical knowledge, while highly knowledgeable students experienced a negative effect of this training approach. Moreover, students' perceptions revealed that facilitative-trained students reported more deep-level learning, while the directive training approach turned out to score higher in terms of quality and perceived effects.

  16. A combined theoretical and in vitro modeling approach for predicting the magnetic capture and retention of magnetic nanoparticles in vivo

    Science.gov (United States)

    David, Allan E.; Cole, Adam J.; Chertok, Beata; Park, Yoon Shin; Yang, Victor C.

    2011-01-01

    Magnetic nanoparticles (MNP) continue to draw considerable attention as potential diagnostic and therapeutic tools in the fight against cancer. Although many interacting forces present themselves during magnetic targeting of MNP to tumors, most theoretical considerations of this process ignore all except for the magnetic and drag forces. Our validation of a simple in vitro model against in vivo data, and subsequent reproduction of the in vitro results with a theoretical model indicated that these two forces do indeed dominate the magnetic capture of MNP. However, because nanoparticles can be subject to aggregation, and large MNP experience an increased magnetic force, the effects of surface forces on MNP stability cannot be ignored. We accounted for the aggregating surface forces simply by measuring the size of MNP retained from flow by magnetic fields, and utilized this size in the mathematical model. This presumably accounted for all particle-particle interactions, including those between magnetic dipoles. Thus, our “corrected” mathematical model provided a reasonable estimate of not only fractional MNP retention, but also predicted the regions of accumulation in a simulated capillary. Furthermore, the model was also utilized to calculate the effects of MNP size and spatial location, relative to the magnet, on targeting of MNPs to tumors. This combination of an in vitro model with a theoretical model could potentially assist with parametric evaluations of magnetic targeting, and enable rapid enhancement and optimization of magnetic targeting methodologies. PMID:21295085

  17. Testing strategies for embryo-fetal toxicity of human pharmaceuticals. Animal models vs. in vitro approaches: a workshop report.

    Science.gov (United States)

    van der Laan, Jan Willem; Chapin, Robert E; Haenen, Bert; Jacobs, Abigail C; Piersma, Aldert

    2012-06-01

    Reproductive toxicity testing is characterized by high animal use. For registration of pharmaceutical compounds, developmental toxicity studies are usually conducted in both rat and rabbits. Efforts have been underway for a long time to design alternatives to animal use. Implementation has lagged, partly because of uncertainties about the applicability domain of the alternatives. The reproductive cycle is complex and not all mechanisms of development can be mimicked in vitro. Therefore, efforts are underway to characterize the available alternative tests with regard to the mechanism of action they include. One alternative test is the mouse embryonic stem cell test (EST), which has been studied since the late 1990s. It is a genuine 3R "alternative" assay as it is essentially animal-free. A meeting was held to review the state-of-the-art of various in vitro models for prediction of developmental toxicity. Although the predictivity of individual assays is improving, a battery of several assays is likely to have even higher predictivity, which is necessary for regulatory acceptance. The workshop concluded that an important first step is a thorough survey of the existing rat and rabbit studies, to fully characterize the frequency of responses and the types of effects seen. At the same time, it is important to continue the optimization of in vitro assays. As more experience accumulates, the optimal conditions, assay structure, and applicability of the alternative assays are expected to emerge. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. Influence of grinding on the nutritive value of peas for ruminants: comparison between in vitro and in situ approaches.

    Science.gov (United States)

    Giger-Reverdin, Sylvie; Maaroufi, Chiraze; Chapoutot, Patrick; Peyronnet, Corinne; Sauvant, Daniel

    2014-07-01

    In ruminant nutrition, peas are characterized by high protein solubility and degradability, which impair its protein value estimated by the official in situ method. Grinding can be used as a technological treatment of pea seeds to modify their nutritional value. The aim of this study was to compare the in situ method with an in vitro method on the same pea either in a coarse pea flour form (PCF) or in a ground pea fine flour form (PFF) to understand the effect of grinding. Both forms were also reground (GPCF and GPFF). PCF presented a lower rate of in vitro degradation than PFF, and more stable fermentation parameters (pH, ammonia, soluble carbohydrates) even if gas production was higher for the PCF after 48 h of incubation. In situ dry matter and protein degradation were lower for PCF than those for PFF; these differences were more marked than with the in vitro method. Reground peas were very similar to PFF. The values for pea protein digestible in the intestine (PDI) were higher for PCF than those for PFF. This study points out the high sensitivity of the in situ method to grinding. The study needs to be validated by in vivo measurements.

  19. Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells

    DEFF Research Database (Denmark)

    Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin

    2016-01-01

    In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured....... In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs) was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three......-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material...

  20. Microscale In Vitro Assays for the Investigation of Neutral Red Retention and Ethoxyresorufin-O-Deethylase of Biofuels and Fossil Fuels

    DEFF Research Database (Denmark)

    Heger, Sebastian; Bluhm, Kerstin; Brendt, Julia

    2016-01-01

    Only few information on the potential toxic effectiveness of biofuels are available. Due to increasing worldwide demand for energy and fuels during the past decades, biofuels are considered as a promising alternative for fossil fuels in the transport sector. Hence, more information on their hazard......, ecotoxicological methods are applied to gain information on potential adverse environmental effects of biofuels at an early phase of their development. In the present study, three potential biofuels, ethyl levulinate, 2-methyltetrahydrofuran and 2-methylfuran were tested. Furthermore, we investigated a fossil...... gasoline fuel, a fossil diesel fuel and an established biodiesel. Two in vitro bioassays, one for assessing cytotoxicity and one for aryl hydrocarbon receptor agonism, so called dioxin-like activity, as measured by Ethoxyresorufin-O-Deethylase, were applied using the permanent fish liver cell line RTL-W1...

  1. Investigation on Secondary Structure Perturbations of Proteins Embedded in Solid Lipid Matrices as a Novel Indicator of their Biological Activity upon In Vitro Release

    DEFF Research Database (Denmark)

    Zeeshan, Farrukh; Tabbassum, Misbah; Jorgensen, Lene

    2018-01-01

    Protein biologics are prone to conformational changes during formulation development. Limited methods are available for conformational analysis of proteins in solid state and in the presences of formulation excipients. The aim of this study was to investigate the secondary structures of proteins...... encased in solid lipid matrices as a novel indicator of their stability upon in vitro release. Model proteins namely catalase and lysozyme were incorporated into lipid namely Precirol® AT05 (glycerol palmitostearate, melting point 58°C) at 30% w/w loading using melting and mixing and wet granulation...... methods. Attenuated total reflectance (ATR-FTIR) spectroscopy, size-exclusion chromatography (SEC) and biological activity analyses were performed. The information about secondary structure was acquired using second derivative analysis of amide-I band (1600-1700 cm(-1)). ATR analysis demonstrated...

  2. A Micro-Level Event-Centered Approach to Investigating Armed Conflict and Population Responses

    OpenAIRE

    Williams, Nathalie E.; Ghimire, Dirgha J.; Axinn, William G.; Jennings, Elyse A.; Pradhan, Meeta S.

    2012-01-01

    In this article, we construct and test a micro-level event-centered approach to the study of armed conflict and behavioral responses in the general population. Event-centered approaches have been successfully used in the macro-political study of armed conflict but have not yet been adopted in micro-behavioral studies. The micro-level event-centered approach that we advocate here includes decomposition of a conflict into discrete political and violent events, examination of the mechanisms thro...

  3. Investigating the relationships between approaches to learning, learner identities and academic achievement in higher education

    DEFF Research Database (Denmark)

    Herrmann, Kim Jesper; Bager-Elsborg, Anna; McCune, Velda

    2017-01-01

    This paper considers relationships between approaches to learning, learner identities, self-efficacy beliefs and academic achievement in higher education. In addition to already established survey instruments, a new scale, subject area affinity, was developed. The scale explores the extent to which....... The new scale also correlated negatively with the surface approach scale. K-means cluster analysis identified seven distinct groups of students who espoused interpretable combinations of approaches, self-efficacy and subject area affinity. Cluster membership was associated with differences in academic...

  4. An in vitro investigation of a newer intracanal medicament Nisin on Enterococcus faecalis in comparison with chlorhexidine and calcium hydroxide

    Directory of Open Access Journals (Sweden)

    Suneel Kumar Chinni

    2011-01-01

    Full Text Available Aim: To determine the antibacterial efficacy of Nisin in comparison with Calcium hydroxide and Chlorhexidine. Materials and Methods: Fifty extracted lower premolar single-rooted teeth were collected and were instrumented with K3 rotary files. Then, root canals were inoculated with a bacterial solution of Enterococcus faecalis. After 21 days, the canals were inoculated with Saline, Nisin, Vancomycin, Calcium hydroxide, and Chlorhexidine. The roots were left for 7 days and on the 8 th day, to investigate the degree of infection of the radicular dentin, specimens of the dentin chips from the full length of the root canal were harvested using a sterile rotary K3 instrument (size 25 6% taper. Results: The results of the present study showed that Nisin and Chlorhexidine showed none of Colony Forming Units (CFU in their respective group. Conclusion: Within the limits of the study, Nisin was effective in eradicating E. faecalis cells in pure culture and root canal dentin.

  5. Investigation of hemodynamics in an in vitro system simulating left ventricular support through the right subclavian artery using 4-dimensional flow magnetic resonance imaging.

    Science.gov (United States)

    Jung, Bernd; Müller, Christoph; Buchenberg, Waltraud; Ith, Michael; Reineke, David; Beyersdorf, Friedhelm; Benk, Christoph

    2015-07-01

    Left ventricular assist devices are an important treatment option for patients with heart failure alter the hemodynamics in the heart and great vessels. Because in vivo magnetic resonance studies of patients with ventricular assist devices are not possible, in vitro models represent an important tool to investigate flow alterations caused by these systems. By using an in vitro magnetic resonance-compatible model that mimics physiologic conditions as close as possible, this work investigated the flow characteristics using 4-dimensional flow-sensitive magnetic resonance imaging of a left ventricular assist device with outflow via the right subclavian artery as commonly used in cardiothoracic surgery in the recent past. An in vitro model was developed consisting of an aorta with its supra-aortic branches connected to a left ventricular assist device simulating the pulsatile flow of the native failing heart. A second left ventricular assist device supplied the aorta with continuous flow via the right subclavian artery. Four-dimensional flow-sensitive magnetic resonance imaging was performed for different flow rates of the left ventricular assist device simulating the native heart and the left ventricular assist device providing the continuous flow. Flow characteristics were qualitatively and quantitatively evaluated in the entire vessel system. Flow characteristics inside the aorta and its upper branching vessels revealed that the right subclavian artery and the right carotid artery were solely supported by the continuous-flow left ventricular assist device for all flow rates. The flow rates in the brain-supplying arteries are only marginally affected by different operating conditions. The qualitative analysis revealed only minor effects on the flow characteristics, such as weakly pronounced vortex flow caused by the retrograde flow via the brachiocephalic artery. The results indicate that, despite the massive alterations in natural hemodynamics due to the

  6. A preliminary investigation into the impact of a pesticide combination on human neuronal and glial cell lines in vitro.

    Directory of Open Access Journals (Sweden)

    Michael D Coleman

    Full Text Available Many pesticides are used increasingly in combinations during crop protection and their stability ensures the presence of such combinations in foodstuffs. The effects of three fungicides, pyrimethanil, cyprodinil and fludioxonil, were investigated together and separately on U251 and SH-SY5Y cells, which can be representative of human CNS glial and neuronal cells respectively. Over 48h, all three agents showed significant reductions in cellular ATP, at concentrations that were more than tenfold lower than those which significantly impaired cellular viability. The effects on energy metabolism were reflected in their marked toxic effects on mitochondrial membrane potential. In addition, evidence of oxidative stress was seen in terms of a fall in cellular thiols coupled with increases in the expression of enzymes associated with reactive species formation, such as GSH peroxidase and superoxide dismutase. The glial cell line showed significant responsiveness to the toxin challenge in terms of changes in antioxidant gene expression, although the neuronal SH-SY5Y line exhibited greater vulnerability to toxicity, which was reflected in significant increases in caspase-3 expression, which is indicative of the initiation of apoptosis. Cyprodinil was the most toxic agent individually, although oxidative stress-related enzyme gene expression increases appeared to demonstrate some degree of synergy in the presence of the combination of agents. This report suggests that the impact of some pesticides, both individually and in combinations, merits further study in terms of their impact on human cellular health.

  7. Evaluation of the antidiabetic property of aqueous leaves extract of Zanthoxylum armatum DC. using in vivo and in vitro approaches

    Directory of Open Access Journals (Sweden)

    Carey Vana Rynjah

    2018-01-01

    Full Text Available The present study was designed to evaluate the antidiabetic potential of the aqueous leaves extract of Zanthoxylum armatum DC. leaves using in vivo and in vitro approaches. For in vivo studies, blood glucose level was monitored at different intervals after administration of varying doses of the extract for its hypoglycemic (100–6000 mg/kg b.w. and antihyperglycemic (250 mg/kg b.w. effect in normoglycemic and diabetic mice. In vitro enzymatic inhibition activity was tested against α-amylase, α- and β-glucosidase and lipase. Additionally hydroxyl radical, hydrogen peroxide scavenging assay and phytochemical screening were also performed. Element analysis of the plant was studied by Atomic Absorption Spectrometry (AAS and Inductively Coupled Plasma Atomic Emission Spectrometer (ICP-AES. The plant extract showed significant hypoglycemic and antihyperglycemic effect in normoglycemic and diabetic mice. The IC50 values of extract for α-amylase, β-glucosidase, lipase, hydroxyl radical scavenging activity, hydrogen peroxide scavenging activity were 7.40 mg/ml, 0.30 mg/ml, 8.35 mg/ml, 3.25 mg/ml, 9.62 mg/ml respectively and the percentage of inhibition for α-glucosidase was 79.82% at 0.8 mg/ml. In vitro studies were compared with their respective standards. Elemental analysis revealed the presence of essential elements such as Mg, V, Fe, Cr, Zn, Cu, Mo, Mn, K, Ca, P and Sr which are all known to play a role in regulating blood glucose. The results demonstrate that Z. armatum aqueous leaves extract possess antidiabetic property in both in vivo and in vitro condition.

  8. Investigation into drug release from colon-specific azoreductase-activated steroid prodrugs using in-vitro models.

    Science.gov (United States)

    Ruiz, Juan F Marquez; Kedziora, Kinga; Windle, Henry; Kelleher, Dermot P; Gilmer, John F

    2011-06-01

    The aim of this study was to investigate drug release from a double steroid prodrug, OPN501, which incorporates a phenylpropionate linker, and its phenylacetate analogue. The prodrugs, which were designed to deliver prednisolone to the colon for the treatment of inflammatory bowel disease, are based on a novel design that requires sequential azoreductase activity and cyclization of an amino ester to trigger drug release. We sought to explain the divergent effects of the two compounds in anti-inflammatory models and to justify the selection of OPN-501 for clinical development. The compounds were incubated in mouse colonic contents (10%) fermented in brain heart infusion under anaerobic conditions. The disappearance of the prodrugs and release of prednisolone was monitored by HPLC. We then developed a method for assessment of prodrug activation using suspensions of Clostridium perfringens, an anaerobe from the human colon. The cyclization of the compounds was studied in various media, assessing the influence of pH and bulk solvent polarity on cyclization rate using HPLC and NMR. The prodrugs were activated via multiple pathways releasing prednisolone in mouse colonic ferment. The compounds released prednisolone by reduction-cyclization in C perfringens suspension. The active OPN-501 generated a stoichiometric amount of prednisolone following azoreductase activation, whereas its analogue did not. The pH rate profile for the cyclization of the amino intermediates of the two compounds revealed significant differences in rate at pH values relevant to the inflamed colon, which explain in part the different amounts of drug produced. The steroid prodrug OPN-501 has optimal drug release characteristics for colon targeting because of a kinetic advantage of a six-membered ring formation in the aminolysis reactions of anilides. The results are relevant to the development of OPN-501 but also to cyclization strategies in prodrug design especially for colon targeting. © 2011 The

  9. In vivo and in vitro investigations of retinal fluorophores in age-related macular degeneration by fluorescence lifetime imaging

    Science.gov (United States)

    Hammer, M.; Quick, S.; Klemm, M.; Schenke, S.; Mata, N.; Eitner, A.; Schweitzer, D.

    2009-02-01

    Ocular fundus autofluorescence imaging has been introduced into clinical diagnostics recently for the observation of the age pigment lipofuscin, a precursor of age-related macular degeneration (AMD). However, a deeper understanding of the generation of single compounds contributing to the lipofuscin as well as of the role of other fluorophores such as FAD, glycated proteins, and collagen needs their discrimination by fluorescence lifetime imaging (FLIM). FLIM at the ocular fundus is performed using a scanning laser ophthalmoscope equipped with a picosecond laser source (448nm or 468nm respectively, 100ps, 80 MHz repetition rate) and dual wavelength (490-560nm and 560-7600nm) time-correlated single photon counting. A three-exponential fit of the fluorescence decay revealed associations of decay times to anatomical structures. Disease-related features are identified from alterations in decay times and-amplitudes. The in-vivo investigations in patients were paralleled by experiments in an organ culture of the porcine ocular fundus. Photo-oxidative stress was induced by exposure to blue light (467nm, 0.41 mW/mm2). Subsequent analysis (fluorescence microscopy, HPLC, LC-MS) indicated the accumulation of the pyridinium bis-retinoid A2E and its oxidation products as well as oxidized phospholipids. These compounds contribute to the tissue auto-fluorescence and may play a key role in the pathogenesis of AMD. Thus, FLIM observation at the ocular fundus in vivo enhances our knowledge on the etiology of AMD and may become a diagnostic tool.

  10. Investigating meta-approaches for reconstructing gene networks in a mammalian cellular context.

    Directory of Open Access Journals (Sweden)

    Azree Nazri

    Full Text Available The output of state-of-the-art reverse-engineering methods for biological networks is often based on the fitting of a mathematical model to the data. Typically, different datasets do not give single consistent network predictions but rather an ensemble of inconsistent networks inferred under the same reverse-engineering method that are only consistent with the specific experimentally measured data. Here, we focus on an alternative approach for combining the information contained within such an ensemble of inconsistent gene networks called meta-analysis, to make more accurate predictions and to estimate the reliability of these predictions. We review two existing meta-analysis approaches; the Fisher transformation combined coefficient test (FTCCT and Fisher's inverse combined probability test (FICPT; and compare their performance with five well-known methods, ARACNe, Context Likelihood or Relatedness network (CLR, Maximum Relevance Minimum Redundancy (MRNET, Relevance Network (RN and Bayesian Network (BN. We conducted in-depth numerical ensemble simulations and demonstrated for biological expression data that the meta-analysis approaches consistently outperformed the best gene regulatory network inference (GRNI methods in the literature. Furthermore, the meta-analysis approaches have a low computational complexity. We conclude that the meta-analysis approaches are a powerful tool for integrating different datasets to give more accurate and reliable predictions for biological networks.

  11. A micro-level event-centered approach to investigating armed conflict and population responses.

    Science.gov (United States)

    Williams, Nathalie E; Ghimire, Dirgha J; Axinn, William G; Jennings, Elyse A; Pradhan, Meeta S

    2012-11-01

    In this article, we construct and test a micro-level event-centered approach to the study of armed conflict and behavioral responses in the general population. Event-centered approaches have been successfully used in the macro-political study of armed conflict but have not yet been adopted in micro-behavioral studies. The micro-level event-centered approach that we advocate here includes decomposition of a conflict into discrete political and violent events, examination of the mechanisms through which they affect behavior, and consideration of differential risks within the population. We focus on two mechanisms: instability and threat of harm. We test this approach empirically in the context of the recent decade-long armed conflict in Nepal, using detailed measurements of conflict-related events and a longitudinal study of first migration, first marriage, and first contraceptive use. Results demonstrate that different conflict-related events independently shaped migration, marriage, and childbearing and that they can simultaneously influence behaviors in opposing directions. We find that violent events increased migration, but political events slowed migration. Both violent and political events increased marriage and contraceptive use net of migration. Overall, this micro-level event-centered approach yields a significant advance for the study of how armed conflict affects civilian behavioral responses.

  12. A framework for investigating mobile web success in the context of e-commerce:an analytic network process (ANP) approach

    OpenAIRE

    Salehi, Mona; Keramati, Abbas; Didehkhani, H.

    2010-01-01

    This study proposes a framework to investigate the factors of mobile web success in the context of e-commerce, and the relative importance of these success factors in selecting the most preferred mobile web. First, the Updated Delone and Mclean IS success model (2003) is chosen to extract significant mobile web success factors in the context of e-commerce. Second, it is extended through applying an Analytic Network Process (ANP) approach for investigating therelative importance of each factor...

  13. Investigation of the in vitro toxicological properties of the synthetic cannabimimetic drug CP-47,497-C8

    Energy Technology Data Exchange (ETDEWEB)

    Koller, Verena J., E-mail: verena.koller@meduniwien.ac.at [Institute of Cancer Research, Department of Internal Medicine 1, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8A, 1090 Vienna (Austria); Auwärter, Volker, E-mail: volker.auwaerter@uniklinik-freiburg.de [Institute of Forensic Medicine, University Medical Center Freiburg, Albertstraße 9, 79104 Freiburg (Germany); Grummt, Tamara, E-mail: tamara.grummt@uba.de [German Federal Environmental Agency, Heinrich-Heine-Str., 12, 08645 Bad Elster (Germany); Moosmann, Bjoern, E-mail: bjoern.moosmann@uniklinik-freiburg.de [Institute of Forensic Medicine, University Medical Center Freiburg, Albertstraße 9, 79104 Freiburg (Germany); Mišík, Miroslav, E-mail: miroslav.misik@meduniwien.ac.at [Institute of Cancer Research, Department of Internal Medicine 1, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8A, 1090 Vienna (Austria); Knasmüller, Siegfried, E-mail: siegfried.knasmueller@meduniwien.ac.at [Institute of Cancer Research, Department of Internal Medicine 1, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8A, 1090 Vienna (Austria)

    2014-06-01

    Cannabicyclohexanol (CP-47,497-C8) is a representative of a group of cannabimimetic cyclohexylphenols which is added to herbal mixtures as a cannabis substitute since 2008. Although in the beginning CP-47,497-C8 was the main ingredient of “Spice” and similar products, it was partly replaced by aminoalkylindole-type cannabinoid receptor agonists like JWH-018, JWH-073 or JWH-250, but never completely disappeared from the market. Since information on its toxicological properties is scarce, we investigated the effects of the drug in human derived cell lines. The cytotoxic effects were studied in a panel of assays (SRB, XTT, LDHe and NR tests) in a buccal derived (TR146) and a liver derived (HepG2) cell line. The strongest effects were seen in the two former assays at levels ≥ 7.5 μM indicating that the compound interferes with protein synthesis and causes membrane damage. In additional comet assays, DNA damage was detected at levels ≥ 10 μM. Experiments with lesion specific enzymes showed that these effects are not due to oxidative damage of DNA bases. The negative findings obtained in Salmonella/microsome assays and the positive results of micronucleus tests with the cell lines indicate that the compound does not cause gene mutations but acts on the chromosomal level. In contrast to other synthetic cannabinoids, no indication for estrogenic/antiestrogenic properties was seen in a luciferase assay with bone marrow derived U2-OS cells. In conclusion, our findings show that the drug has only weak cytotoxic properties. However, the induction of chromosomal damage indicates that it may cause adverse effects in users due to its impact on the stability of the genetic material. - Highlights: • We tested the toxic properties of a synthetic cannabinoid. • Acute cytotoxic effects were detected with doses ≥ 7 μM. • No hormonal effects were found. • DNA damage was detected at levels ≥ 10 μM in comet assay and micronucleus tests. • Effects in directly

  14. A qualitative and quantitative investigation into the effect of fluoride formulations on enamel erosion and erosion-abrasion in vitro.

    Science.gov (United States)

    Austin, R S; Stenhagen, K S; Hove, L H; Dunne, S; Moazzez, R; Bartlett, D W; Tveit, A B

    2011-10-01

    To investigate the effect of a single application of highly concentrated SnF(2) and NaF solutions and a NaF/CaF(2) varnish on human enamel subjected to hydrochloric acid erosion and tooth brush abrasion. Forty enamel samples were prepared from human third molars and NaF (9500ppm, pH 8.0), SnF(2) (9500ppm, pH 2.6) solutions; Bifluorid10(®) varnish (42,500ppm, NaF 5%, CaF(2) 5%) and deionized water (control) was applied to the enamel. Following this three, six and nine cycles of erosion [1 cycle=erosion (0.01M HCl, pH 2.2, 2min)+artificial saliva (1h, pH 7.0)] and erosion-abrasion [1 cycle=erosion (0.01M HCl, pH 2.2, 2min)+artificial saliva (1h, pH 7.0)+abrasion (120 linear strokes in artificial saliva from Tepe medium soft brushes 200g loading)] were carried out. The fluoride treated enamel was analysed using Knoop microhardness, scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). For erosion alone, there was significantly less microhardness reduction in the Bifluorid10(®) group after three and six cycles of erosion (P0.05). The EDS analysis showed that only the Bifluorid10(®) group had any detectable fluorine following erosion and erosion-abrasion (0.1wt.% and 0.2wt.% fluorine respectively). The surface fluorine was found to have been removed after erosion and erosion-abrasion for all other surface treatments. Although precipitates were observed after application of the surface treatments, following erosion-abrasion, no visible surface effects from any fluoride preparation remained. Enamel surface precipitates from application NaF, SnF(2) solutions appear to not be able to provide protection against gastric erosion and tooth brush abrasion. The NaF/CaF(2) varnish provided limited protection against erosion but the role for such varnishes in gastric erosion and tooth brush abrasion remains uncertain. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Assessing Finger Joint Biomechanics by Applying Equal Force to Flexor Tendons In Vitro Using a Novel Simultaneous Approach.

    Directory of Open Access Journals (Sweden)

    Tai-Hua Yang

    Full Text Available The flexor digitorum superficialis (FDS and flexor digitorum profundus (FDP are critical for finger flexion. Although research has recently focused on these tendons' coactivity, their contributions in different tasks remain unclear. This study created a novel simultaneous approach to investigate the coactivity between the tendons and to clarify their contributions in different tasks.Ten human cadaveric hands were mounted on our custom frame with the FDS and FDP of the third finger looped through a mechanical pulley connected to a force transducer. Joint range of motion, tendon excursion and loading force were recorded during individual joint motion and free joint movement from rest to maximal flexion. Each flexor tendon's moment arm was then calculated.In individual motions, we found that the FDP contributed more than the FDS in proximal interphalangeal (PIP joint motion, with an overall slope of 1.34 and all FDP-to-FDS excursion (P/S ratios greater than 1.0 with force increase. However, the FDP contributed less than the FDS in metacarpophalangeal (MCP joint motion, with an overall slope of 0.95 and P/S ratios smaller than 1.0 throughout the whole motion except between 1.9% and 13.1% force. In free joint movement, the FDP played a greater role than the FDS, with an overall ratio of 1.37 and all P/S ratios greater than 1.0.The new findings include differences in finger performance and excursion amounts between the FDS and FDP throughout flexion. Such findings may provide the basis for new hand models and treatments.

  16. New approach for a reliable in vitro sun protection factor method - Part II: Practical aspects and implementations.

    Science.gov (United States)

    Miksa, S; Lutz, D; Guy, C; Delamour, E

    2016-10-01

    Our previous paper (Part I: Principle and mathematical aspects) presented a new reliable in vitro sun protection factor (SPF) method and demonstrated it to be reproducible and correlated with the in vivo method. Nevertheless, the relevance of an international method should to be adaptable to all products on the market and demonstrated with a blind test. Thus, the aim of this second article was to focus on the practical aspects and implementation (Part II) of a large population of different commercially available sunscreen formulations to obtain similar in vivo SPF results for the purpose of labelling. The method uses the spectroradiometric measurement of residual ultraviolet (UV) through the sample that was applied on a substrate with a robotic appliance. The method has been demonstrated to be highly reliable, and it is based on a multisubstrate solution with a single UV pre-irradiation dose. Furthermore, different categories of the product were studied to identify a reliable and universal in vitro SPF method. Based on different sunscreens products classified into 5 different groups (emulsion, oil, alcohol, stick and powder), it was demonstrated that our method has good reproducibility and accuracy compared with the clinical SPF method. Indeed, the mean coefficient of variation (CV%) was approximately 7%, and the coefficient of correlation reached approximately 0.8-1.0 for different types of tested products. Our second paper concludes that the new in vitro SPF method (based on 113 sunscreen products from the Parts I and II) is clearly adaptable for the SPF labelling purpose on any product type because it is non-invasive, less expensive, more practical and more reliable if performed under strict conditions. © 2016 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  17. University students' learning approaches in three cultures: an investigation of Biggs's 3P model.

    Science.gov (United States)

    Zhang, L F

    2000-01-01

    The relationship of various learning approaches to students' academic achievement, abilities, and other characteristics was examined in a sample of university students in Hong Kong, mainland China, and the United States. The theoretical framework for this project was J. B. Biggs's (1987) theory of student learning approaches. The participants completed the Study Process Questionnaire (based on Biggs's theory) and provided a variety of demographic information. The participants' achievement scores and self-rated scores on analytical, creative, and practical abilities were also obtained. Results indicated that scores on certain subscales of the Study Process Questionnaire statistically predicted participants' achievement beyond their self-rated abilities. In addition, certain learning approaches were significantly related to the participants' ages, gender, parents' education levels, and their travel and work experiences. Implications of these findings are discussed as they relate to teaching and learning.

  18. Financial Investigations. A Financial Approach to Detecting and Resolving Crimes. [Text], Instructor's Guide, and Student Workbook.

    Science.gov (United States)

    Internal Revenue Service (Dept. of Treasury), Washington, DC.

    This packet contains a textbook, an instructor's guide, and a student workbook for a course on conducting financial investigations to detect and solve crimes. The topics covered in the 11 chapters of the textbook and the ancillaries are the following: (1) why financial investigation?; (2) laws related to financial crimes; (3) evidence; (4) sources…

  19. Investigation of tt in the full hadronic final state at CDF with a neural network approach

    CERN Document Server

    Sidoti, A; Busetto, G; Castro, A; Dusini, S; Lazzizzera, I; Wyss, J

    2001-01-01

    In this work we present the results of a neural network (NN) approach to the measurement of the tt production cross-section and top mass in the all-hadronic channel, analyzing data collected at the Collider Detector at Fermilab (CDF) experiment. We have used a hardware implementation of a feedforward neural network, TOTEM, the product of a collaboration of INFN (Istituto Nazionale Fisica Nucleare)-IRST (Istituto per la Ricerca Scientifica e Tecnologica)-University of Trento, Italy. Particular attention has been paid to the evaluation of the systematics specifically related to the NN approach. The results are consistent with those obtained at CDF by conventional data selection techniques. (38 refs).

  20. Combined In Silico and In Vitro Approach Predicts Low Wall Shear Stress Regions in a Hemofilter that Correlate with Thrombus Formation In Vivo.

    Science.gov (United States)

    Buck, Amanda K W; Groszek, Joseph J; Colvin, Daniel C; Keller, Sara B; Kensinger, Clark; Forbes, Rachel; Karp, Seth; Williams, Phillip; Roy, Shuvo; Fissell, William H

    2017-08-29

    A major challenge in developing blood-contacting medical devices is mitigating thrombogenicity of an intravascular device. Thrombi may interfere with device function or embolize from the device to occlude distant vascular beds with catastrophic consequences. Chemical interactions between plasma proteins and bioengineered surface occur at the nanometer scale; however, continuum models of blood predict local shear stresses that lead to platelet activation or aggregation and thrombosis. Here, an iterative approach to blood flow path design incorporating in silico, in vitro, and in vivo experiments predicted the occurrence and location of thrombi in an implantable hemofilter. Low wall shear stress (WSS) regions identified by computational fluid dynamics (CFD) predicted clot formation in vivo. Revised designs based on CFD demonstrated superior performance, illustrating the importance of a multipronged approach for a successful design process.

  1. A comprehensive approach to qualify and validate the essential parameters of an in vitro release test (IVRT) method for acyclovir cream, 5.

    Science.gov (United States)

    Tiffner, Katrin I; Kanfer, Isadore; Augustin, Thomas; Raml, Reingard; Raney, Sam G; Sinner, Frank

    2017-09-19

    The rate of release of an active pharmaceutical ingredient (API) from a topical semisolid dosage form can be influenced by its physical and structural properties. An In Vitro Release Test (IVRT) is an established method to characterize this rate of API release and compare the underlying sameness in product quality characteristics. The purpose of this work was to validate an IVRT method to compare acyclovir cream, 5% products. However, despite widespread use of the IVRT since 1997, there has been no established approach to validate an IVRT method. Our approach included: 1) qualification of the diffusion cell apparatus, 2) qualification of the laboratory, 3) validation of the HPLC analytical method, and 4) validation of numerous critical parameters of the IVRT method, itself, and resulted in a comprehensive and successful IVRT method validation. Subsequent to the IVRT validation work described here, the U.S. Food and Drug Administration (FDA) drafted a guidance on the development and validation of an IVRT method for acyclovir cream, 5%. Although there are notable differences between our approach and the approach in that guidance, this report illustrates how many of the same essential qualification parameters and validation concepts were considered and systematically addressed in our approach to IVRT validation. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Investigating the Effectiveness of Storylines Embedded within a Context-Based Approach: The Case for the Periodic Table

    Science.gov (United States)

    Demircioglu, Hulya; Demircioglu, Gokhan; Calik, Muammar

    2009-01-01

    We investigated the effect of the context-based approach on 9th grade students' conceptions of the Periodic Table. Within a nonequivalent pretest-posttest control group design the study was conducted with 80 grade 9 students (aged 15-16) drawn from two classes (39 and 41 students) in a high school in Turkey. The experimental group was exposed to…

  3. Mastery-Approach Goals and Knowledge Transfer: An Investigation into the Effects of Task Structure and Framing Instructions

    Science.gov (United States)

    Belenky, Daniel M.; Nokes-Malach, Timothy J.

    2013-01-01

    Although prior work has shown that mastery-approach achievement goals are related to positive learning behaviors (e.g., more interest, perseverance, and self-regulation), less is known about how these goals interact with instruction to influence knowledge transfer. To address these issues we conducted a laboratory experiment investigating how two…

  4. Comparison of the Predictability of Human Hepatic Clearance for Organic Anion Transporting Polypeptide Substrate Drugs Between Different In Vitro-In Vivo Extrapolation Approaches.

    Science.gov (United States)

    Izumi, Saki; Nozaki, Yoshitane; Komori, Takafumi; Takenaka, Osamu; Maeda, Kazuya; Kusuhara, Hiroyuki; Sugiyama, Yuichi

    2017-09-01

    Prediction of human pharmacokinetic profiles of drug candidates is an essential step toward first-in-human studies. However, it remains difficult to quantitatively predict hepatic clearance, particularly when hepatic uptake is mediated by transporter(s). Using 15 organic anion transporting polypeptide (OATP) substrate drugs, we tested 3 in vitro-in vivo extrapolation (IVIVE) approaches to predict overall hepatic intrinsic clearance in vivo (CLint,all,vivo). IVIVE approaches involved metabolic intrinsic clearance in human liver microsomes (CLint,met) with or without hepatocyte-to-buffer maximum unbound concentration ratio (Kp,uu,max) correction and uptake intrinsic clearance at 37°C (PSinf,37°C) in human hepatocyte suspensions. Kp,uu,max and PSinf,37°C values were determined in 2 hepatocyte batches, and all tested compounds showed temperature-dependent uptake, consistent with the fact of transporter-mediated uptake. CLint,met substantially underestimated CLint,all,vivo. By multiplying CLint,met by Kp,uu,max values, the prediction performance was much improved; however, in vitro-in vivo correlation was poor. Among the IVIVE approaches, PSinf,37°C showed the most robust correlation with CLint,all,vivo, and one of the hepatocyte batches could predict CLint,all,vivo with a minimal empirical scaling factor. These results suggested IVIVE with hepatic uptake clearance determined in hepatocyte suspensions as one of the most practical approaches for predicting CLint,all,vivo of OATP substrate drugs. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  5. Integrative approaches to investigating human-natural systems: the Baltimore ecosystem study

    Science.gov (United States)

    Mary L. Cadenasso; Steward T.A. Pickett; Morgan J. Grove; Morgan J. Grove

    2006-01-01

    This paper presents an overview of the research approaches used to study metropolitan Baltimore (Maryland, USA) as an ecological system. The urban ecosystem is a complex of biophysical, social, and built components, and is studied by an interdisciplinary teamof biological, social, and physical scientists, and urban designers. Ecology ?of? themetropolis is addressed...

  6. Is Combining Child Labour and School Education the Right Approach? Investigating the Cambodian Case

    Science.gov (United States)

    Kim, Chae-Young

    2009-01-01

    The paper considers whether letting children combine work and school is a valid and effective approach in Cambodia. Policy makers' suggestions that child labour should be allowed to some extent due to household poverty appear ungrounded as no significant relation between children's work and household poverty is found while arranging school…

  7. Investigating the management of diabetes in nursing homes using a mixed methods approach.

    LENUS (Irish Health Repository)

    Hurley, L

    2017-03-21

    As populations age there is an increased demand for nursing home (NH) care and a parallel increase in the prevalence of diabetes. Despite this, there is growing evidence that the management of diabetes in NHs is suboptimal. The reasons for this are complex and poorly understood. This study aimed to identify the current level of diabetes care in NHs using a mixed methods approach.

  8. Investigating the Relationship among Extracurricular Activities, Learning Approach and Academic Outcomes: A Case Study

    Science.gov (United States)

    Chan, Yiu-Kong

    2016-01-01

    Learning effectiveness requires an understanding of the relationship among extracurricular activities, learning approach and academic performance and, it is argued, this helps educators develop techniques designed to enrich learning effectiveness. Biggs' Presage-Process-Product model on student learning has identified the relationship among…

  9. Investigating the Interrelationships among Conceptions of, Approaches to, and Self-Efficacy in Learning Science

    Science.gov (United States)

    Zheng, Lanqin; Dong, Yan; Huang, Ronghuai; Chang, Chun-Yen; Bhagat, Kaushal Kumar

    2018-01-01

    The purpose of this study was to examine the relations between primary school students' conceptions of, approaches to, and self-efficacy in learning science in Mainland China. A total of 1049 primary school students from Mainland China participated in this study. Three instruments were adapted to measure students' conceptions of learning science,…

  10. Approaching Knowledge Management through the Lens of the Knowledge Life Cycle: A Case Study Investigation

    Science.gov (United States)

    Fowlin, Julaine M.; Cennamo, Katherine S.

    2017-01-01

    More organizational leaders are recognizing that their greatest competitive advantage is the knowledge base of their employees and for organizations to thrive knowledge management (KM) systems need to be in place that encourage the natural interplay and flow of tacit and explicit knowledge. Approaching KM through the lens of the knowledge life…

  11. Investigation of the Influence of the As-Grown ZnO Nanorods and Applied Potentials on an Electrochemical Sensor for In-Vitro Glucose Monitoring

    Directory of Open Access Journals (Sweden)

    Mohammed Marie

    2017-01-01

    Full Text Available The influence of the as-grown zinc oxide nanorods (ZnO NRs on the fabricated electrochemical sensor for in vitro glucose monitoring were investigated. A direct growth of ZnO NRs was performed on the Si/SiO2/Au electrode, using hydrothermal and sol-gel techniques at low temperatures. The structure, consisting of a Si/SiO2/Au/GOx/Nafion membrane, was considered as a baseline, and it was tested under several applied potential 0.1–0.8 V. The immobilized working electrode, with GOx and a nafion membrane, was characterized amperometrically using a source meter Keithely 2410, and an electrochemical impedance Gamry potentiostat. The sensor exhibited the following: a high sensitivity of ~0.468 mA/cm2 mM, a low detection limit in the order of 166.6 µM, and a fast and sharp response time of around 2 s. The highest sensitivity and the lowest limit of detection were obtained at 0.4 volt, after the growth of ZnO NRs. The highest net sensitivity was obtained after subtracting the sensitivity of the baseline, and it was in the order of 0.315 mA/cm2·mM. The device was tested with a range of glucose concentrations from 1–10 mM, showing a linear line from 3–8 mM, and the device was saturated after exceeding high concentrations of glucose. Such devices can be used for in vitro glucose monitoring, since glucose changes can be accurately detected.

  12. In Vivo and In Vitro Genotoxic and Epigenetic Effects of Two Types of Cola Beverages and Caffeine: A Multiassay Approach

    Science.gov (United States)

    Merinas-Amo, Tania; Moreno-Millán, Miguel; Alonso-Moraga, Ángeles; Demyda-Peyrás, Sebastián

    2016-01-01

    The aim of this work was to assess the biological and food safety of two different beverages: Classic Coca Cola™ (CCC) and Caffeine-Free Coca Cola (CFCC). To this end, we determined the genotoxicological and biological effects of different doses of lyophilised CCC and CFCC and Caffeine (CAF), the main distinctive constituent. Their toxic/antitoxic, genotoxic/antigenotoxic, and chronic toxicity (lifespan assay) effects were determined in vivo using the Drosophila model. Their cytotoxic activities were determined using the HL-60 in vitro cancer model. In addition, clastogenic DNA toxicity was measured using internucleosomal fragmentation and SCGE assays. Their epigenetic effects were assessed on the HL-60 methylation status using some repetitive elements. The experimental results showed a slight chemopreventive effect of the two cola beverages against HL-60 leukaemia cells, probably mediated by nonapoptotic mechanisms. Finally, CCC and CAF induced a global genome hypomethylation evaluated in LINE-1 and Alu M1 repetitive elements. Overall, we demonstrated for the first time the safety of this famous beverage in in vivo and in vitro models. PMID:27471731

  13. In Vivo and In Vitro Genotoxic and Epigenetic Effects of Two Types of Cola Beverages and Caffeine: A Multiassay Approach

    Directory of Open Access Journals (Sweden)

    Marcos Mateo-Fernández

    2016-01-01

    Full Text Available The aim of this work was to assess the biological and food safety of two different beverages: Classic Coca Cola™ (CCC and Caffeine-Free Coca Cola (CFCC. To this end, we determined the genotoxicological and biological effects of different doses of lyophilised CCC and CFCC and Caffeine (CAF, the main distinctive constituent. Their toxic/antitoxic, genotoxic/antigenotoxic, and chronic toxicity (lifespan assay effects were determined in vivo using the Drosophila model. Their cytotoxic activities were determined using the HL-60 in vitro cancer model. In addition, clastogenic DNA toxicity was measured using internucleosomal fragmentation and SCGE assays. Their epigenetic effects were assessed on the HL-60 methylation status using some repetitive elements. The experimental results showed a slight chemopreventive effect of the two cola beverages against HL-60 leukaemia cells, probably mediated by nonapoptotic mechanisms. Finally, CCC and CAF induced a global genome hypomethylation evaluated in LINE-1 and Alu M1 repetitive elements. Overall, we demonstrated for the first time the safety of this famous beverage in in vivo and in vitro models.

  14. Allicin functionalized locust bean gum nanoparticles for improved therapeutic efficacy: An in silico, in vitro and in vivo approach.

    Science.gov (United States)

    Soumya, R S; Sherin, S; Raghu, K G; Abraham, Annie

    2018-04-01

    The field of nanotechnology has overgrown over the past few years and has even ventured into the field of medicine. The aim of the present study is to develop a novel allicin functionalized locust bean gum nanoparticle using the nanoprecipitation technique. The synthesized nanoparticles were characterized by dynamic light scattering, scanning electron microscopy and transmission electron microscopy. The characterization study revealed the nanoscale structure (∼100nm) of the prepared particles. In silico toxicology analysis were carried out to assess the drug-like properties and virtual toxicity of allicin. Toxicity of the prepared nanoparticles were carried out in RAW 264.7 cell lines in vitro and in vivo studies were carried out in Sprague-Dawley rats. In in vitro study, LBGAN showed a maximum toxicity of 10.51% in MTT assay, no reactive oxygen species generation on DCFDA staining and LBGAN was effective to protect the cells from apoptosis. In in vivo toxicity studies LBGAN showed no significant change in the activities of the marker enzymes like LDH, CK-MB, ALP, ACP, AST and ALT. Thus, the functionalization of nanoparticles with allicin has the benefit of providing protection and stability to the allicin, in addition to increasing its pharmacological activity. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Statistical optimization and in vitro evaluation of metformin hydrochloride asymmetric membrane capsules prepared by a novel semiautomatic manufacturing approach.

    Science.gov (United States)

    Banala, Venkatesh Teja; Srinivasan, Bharath; Rajamanickam, Deveswaran; Basappa Veerbadraiah, Basavaraj; Varadarajan, Madhavan

    2013-01-01

    Asymmetric membrane capsules (AMCs) are one of the novel osmotic delivery devices which deliver a wide range of drugs in controlled manner. In the present work, we developed and validated a semiautomatic process by fabricating a hydraulic assisted bench top model for manufacturing AMCs. The capsule walls of AMCs were prepared by dip coating phase inversion process using cellulose acetate butyrate (CAB) as coating polymer and propylene glycol (PG) as plasticizer and pore former. The comparative examination of physical parameters confirmed the consistency, efficiency, and reproducibility of the semiautomatic process over the manual procedure. The SEM studies revealed a thin dense region supported on a thicker porous membrane of the capsule shells. Formulations of AMCs were prepared based on a 2(3) full factorial design using metformin hydrochloride as the model drug. The effect of formulation variables such as concentration of PG and levels of fructose and potassium chloride were studied on the in vitro drug release using Design-Expert 8.0.2 (USA) software. From the in vitro release studies, it was observed that the concentration of pore former and level of osmogents had a direct effect on the drug release. From the validation studies of the optimized formulation (OPT) with the predicted response, it was observed that the drug release was independent of pH and agitation intensity but dependent on osmotic pressure of the dissolution medium. The OPT followed controlled zero-order release kinetics over a period of 13 h.

  16. A Case Investigation of Product Structure Complexity in Mass Customization Using a Data Mining Approach

    DEFF Research Database (Denmark)

    Nielsen, Peter; Brunø, Thomas Ditlev; Nielsen, Kjeld

    2014-01-01

    This paper presents a data mining method for analyzing historical configuration data providing a number of opportunities for improving mass customization capabilities. The overall objective of this paper is to investigate how specific quantitative analyses, more specifically the association rule...

  17. An experimental approach for investigating consumers evaluation of pharmacist consultation services

    OpenAIRE

    Schommer, Jon C.

    2000-01-01

    The goal of this study was to investigate factors that influence consumers’ perceptions of service encounter satisfaction, overall service quality, and trust in the service provider for pharmacist consultation services. We used the Dynamic Process Model of Service Quality as the framework for investigating the formation of these evaluations. Consumers’ prior expectations of what should and will transpire during the service episode(s) and the performance level of the actual delivered service d...

  18. In vitro approaches to assess the effects of açai (Euterpe oleracea) digestion on polyphenol availability and the subsequent impact on the faecal microbiota

    OpenAIRE

    Alqurashi, Randah M.; Alarifi, Sehad N.; Walton, Gemma E.; Costabile, Adele F.; Rowland, Ian R.; Commane, Daniel M.

    2017-01-01

    A considerable proportion of dietary plant-polyphenols reach the colon intact; determining the effects of these compounds on colon-health is of interest. We hypothesise that both fibre and plant polyphenols present in açai (Euterpe oleracea) provide prebiotic and anti-genotoxic benefits in the colon. We investigated this hypothesis using a simulated in vitro gastrointestinal digestion of açai pulp, and a subsequent pH-controlled, anaerobic, batch-culture fermentation model reflective of the d...

  19. A Tox21 Approach to Altered Epigenetic Landscapes: Assessing Epigenetic Toxicity Pathways Leading to Altered Gene Expression and Oncogenic Transformation In Vitro.

    Science.gov (United States)

    Parfett, Craig L; Desaulniers, Daniel

    2017-06-01

    , HOTAIR and ANRIL) were found to have experimental evidence showing that functional perturbations played "driver" roles in human cellular transformation. Measurement of epigenotoxicants presents challenges for short-term carcinogenicity testing, especially in the high-throughput modes emphasized in the Tox21 chemicals testing approach. There is need to develop and validate in vitro tests to detect both, locus-specific, and genome-wide, epigenetic alterations with causal links to oncogenic cellular phenotypes. Some recent examples of cell-based high throughput chemical screening assays are presented that have been applied or have shown potential for application to epigenetic endpoints.

  20. A Tox21 Approach to Altered Epigenetic Landscapes: Assessing Epigenetic Toxicity Pathways Leading to Altered Gene Expression and Oncogenic Transformation In Vitro

    Science.gov (United States)

    Parfett, Craig L.; Desaulniers, Daniel

    2017-01-01

    , HOTAIR and ANRIL) were found to have experimental evidence showing that functional perturbations played “driver” roles in human cellular transformation. Measurement of epigenotoxicants presents challenges for short-term carcinogenicity testing, especially in the high-throughput modes emphasized in the Tox21 chemicals testing approach. There is need to develop and validate in vitro tests to detect both, locus-specific, and genome-wide, epigenetic alterations with causal links to oncogenic cellular phenotypes. Some recent examples of cell-based high throughput chemical screening assays are presented that have been applied or have shown potential for application to epigenetic endpoints. PMID:28587163

  1. A Tox21 Approach to Altered Epigenetic Landscapes: Assessing Epigenetic Toxicity Pathways Leading to Altered Gene Expression and Oncogenic Transformation In Vitro

    Directory of Open Access Journals (Sweden)

    Craig L. Parfett

    2017-06-01

    A, UHRF1, CTCF, HOTAIR and ANRIL were found to have experimental evidence showing that functional perturbations played “driver” roles in human cellular transformation. Measurement of epigenotoxicants presents challenges for short-term carcinogenicity testing, especially in the high-throughput modes emphasized in the Tox21 chemicals testing approach. There is need to develop and validate in vitro tests to detect both, locus-specific, and genome-wide, epigenetic alterations with causal links to oncogenic cellular phenotypes. Some recent examples of cell-based high throughput chemical screening assays are presented that have been applied or have shown potential for application to epigenetic endpoints.

  2. Investigation of X-Ray Thomson Scattering Using A Statistical Approach

    Science.gov (United States)

    Johnson, Laura

    2014-10-01

    We present a statistical method of computing x-ray Thomson scattering signals. This model uses average atom wave functions for both bound and continuum electrons, which are computed in a spherically symmetric, self-consistent potential. The wave functions are used to obtain electron distributions for a statistical approach to computing the scattering signals. We compare the differences between using distorted-wave continuum electrons and free-wave electrons in both the statistical approach and the impulse approximation. The results are compared to various experiments including experimental data taken at Cornell's Laboratory of Plasma Studies. Sandia is a multi-program laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy under Contract DE-AC04-94AL85000.

  3. A multiscale approach to blast neurotrauma modeling:Part II: Methodology for inducing blast injury to in vitro models

    Directory of Open Access Journals (Sweden)

    Gwen B. Effgen

    2012-02-01

    Full Text Available Due to the prominent role of improvised explosive devices (IEDs in wounding patterns of U.S. war-fighters in Iraq and Afghanistan, blast injury has risen to a new level of importance and is recognized to be a major cause of injuries to the brain. However, an injury risk-function for microscopic, macroscopic, behavioral, and neurological deficits has yet to be defined. While operational blast injuries can be very complex and thus difficult to analyze, a simplified blast injury model would facilitate studies correlating biological outcomes with blast biomechanics to define tolerance criteria. Blast-induced traumatic brain injury (bTBI results from the translation of a shock wave in air, such as that produced by an IED, into a pressure wave within the skull-brain complex. Our blast injury methodology recapitulates this phenomenon in vitro, allowing for control of the injury biomechanics via a compressed-gas shock tube used in conjunction with a custom-designed, fluid-filled receiver that contains the living culture. The receiver converts the air shock wave into a fast-rising pressure transient with minimal reflections, mimicking the intracranial pressure history in blast. We have developed an organotypic hippocampal slice culture model that exhibits cell death when exposed to a 530  17.7 kPa peak overpressure with a 1.026 ± 0.017 ms duration and 190 ± 10.7 kPa-ms impulse in-air. We have also injured a simplified in vitro model of the blood-brain barrier, which exhibits disrupted integrity immediately following exposure to 581  10.0 kPa peak overpressure with a 1.067 ms ± 0.006 ms duration and 222 ± 6.9 kPa-ms impulse in-air. To better prevent and treat bTBI, both the initiating biomechanics and the ensuing pathobiology must be understood in greater detail. A well-characterized, in vitro model of bTBI, in conjunction with animal models, will be a powerful tool for developing strategies to mitigate the risks of bTBI.

  4. A Multiscale Approach to Blast Neurotrauma Modeling: Part II: Methodology for Inducing Blast Injury to in vitro Models.

    Science.gov (United States)

    Effgen, Gwen B; Hue, Christopher D; Vogel, Edward; Panzer, Matthew B; Meaney, David F; Bass, Cameron R; Morrison, Barclay

    2012-01-01

    Due to the prominent role of improvised explosive devices (IEDs) in wounding patterns of U.S. war-fighters in Iraq and Afghanistan, blast injury has risen to a new level of importance and is recognized to be a major cause of injuries to the brain. However, an injury risk-function for microscopic, macroscopic, behavioral, and neurological deficits has yet to be defined. While operational blast injuries can be very complex and thus difficult to analyze, a simplified blast injury model would facilitate studies correlating biological outcomes with blast biomechanics to define tolerance criteria. Blast-induced traumatic brain injury (bTBI) results from the translation of a shock wave in-air, such as that produced by an IED, into a pressure wave within the skull-brain complex. Our blast injury methodology recapitulates this phenomenon in vitro, allowing for control of the injury biomechanics via a compressed-gas shock tube used in conjunction with a custom-designed, fluid-filled receiver that contains the living culture. The receiver converts the air shock wave into a fast-rising pressure transient with minimal reflections, mimicking the intracranial pressure history in blast. We have developed an organotypic hippocampal slice culture model that exhibits cell death when exposed to a 530 ± 17.7-kPa peak overpressure with a 1.026 ± 0.017-ms duration and 190 ± 10.7 kPa-ms impulse in-air. We have also injured a simplified in vitro model of the blood-brain barrier, which exhibits disrupted integrity immediately following exposure to 581 ± 10.0 kPa peak overpressure with a 1.067 ± 0.006-ms duration and 222 ± 6.9 kPa-ms impulse in-air. To better prevent and treat bTBI, both the initiating biomechanics and the ensuing pathobiology must be understood in greater detail. A well-characterized, in vitro model of bTBI, in conjunction with animal models, will be a powerful tool for developing strategies to mitigate the risks of bTBI.

  5. Using a Comparative Species Approach to Investigate the Neurobiology of Paternal Responses

    OpenAIRE

    Franssen, Catherine L.; Bardi, Massimo; Lambert, Kelly G.

    2011-01-01

    A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors. Using animal models to accomplish this goal, many methodological strategies require invasive techniques to manipulate the intensity of the behavior of interest (e.g., lesion methods, pharmacological manipulations, microdialysis techniques, genetically-engineered animal models). The utilization of a comparative species approach allows researchers to take advantage of naturally ...

  6. HA novel approach to investigate tissue-specific trinucleotide repeat instability

    Directory of Open Access Journals (Sweden)

    Boily Marie-Josee

    2010-03-01

    Full Text Available Abstract Background In Huntington's disease (HD, an expanded CAG repeat produces characteristic striatal neurodegeneration. Interestingly, the HD CAG repeat, whose length determines age at onset, undergoes tissue-specific somatic instability, predominant in the striatum, suggesting that tissue-specific CAG length changes could modify the disease process. Therefore, understanding the mechanisms underlying the tissue specificity of somatic instability may provide novel routes to therapies. However progress in this area has been hampered by the lack of sensitive high-throughput instability quantification methods and global approaches to identify the underlying factors. Results Here we describe a novel approach to gain insight into the factors responsible for the tissue specificity of somatic instability. Using accurate genetic knock-in mouse models of HD, we developed a reliable, high-throughput method to quantify tissue HD CAG repeat instability and integrated this with genome-wide bioinformatic approaches. Using tissue instability quantified in 16 tissues as a phenotype and tissue microarray gene expression as a predictor, we built a mathematical model and identified a gene expression signature that accurately predicted tissue instability. Using the predictive ability of this signature we found that somatic instability was not a consequence of pathogenesis. In support of this, genetic crosses with models of accelerated neuropathology failed to induce somatic instability. In addition, we searched for genes and pathways that correlated with tissue instability. We found that expression levels of DNA repair genes did not explain the tissue specificity of somatic instability. Instead, our data implicate other pathways, particularly cell cycle, metabolism and neurotransmitter pathways, acting in combination to generate tissue-specific patterns of instability. Conclusion Our study clearly demonstrates that multiple tissue factors reflect the level of

  7. Mapping audiovisual translation investigations: research approaches and the role of technology

    OpenAIRE

    Matamala, Anna

    2017-01-01

    This article maps audiovisual translation research by analysing in a contrastive way the abstracts presented at three audiovisual translation conferences ten years ago and nowadays. The comparison deals with the audiovisual transfer modes and topics under discussion, and the approach taken by the authors in their abstracts. The article then shifts the focus to the role of technology in audiovisual translation research, as it is considered an element that is impacting and will continue to impa...

  8. Approach, avoidance and weight-related testing: An investigation of frontal EEG asymmetry.

    Science.gov (United States)

    Faries, Mark D; Kephart, Wesley; Jones, Eric J

    2015-01-01

    Two motivational systems underlie behaviour and affective responses - an inhibition/avoidance system and an activation/approach system. The purpose of the present study was to explore if individual differences in these motivational systems would occur in response to common weight and body composition testing within a sample of young, adult women. Electroencephalogram was used to distinguish approach or avoidance orientations via frontal asymmetry before and after testing sessions. Clear distinctions in motivational response were found, with 65% of the sample responding with an approach motivation, while 35% responded with an avoidance motivation. Even though all participants, on average, experienced a negative affective response, only the avoidance group self-reported a subsequent increase in "comfort food" consumption of desserts and snacks the week following the testing session. As shown with other stressors, clear individual differences exist in motivational responses to common weight and body composition testing. Such testing produces a general negative affective response; however, the individual differences in motivational responses might produce different behavioural choices. Future research and interventions in health communication should be considerate to this variation in motivational responses to help explain changes in both healthy and unhealthy behaviours following interactions involving one's body weight and/or body composition.

  9. Commentary: Utilizing Community-Engaged Approaches to Investigate and Address Hmong Women’s Cancer Disparities

    Directory of Open Access Journals (Sweden)

    Shannon M.A. Sparks

    2014-12-01

    Full Text Available Cancer is a growing concern for women in the Hmong community. Hmong women experience poor health outcomes for both cervical and breast cancer, largely due to low rates of screening and resultant late-stage at diagnosis. Both breast and cervical cancer screening are complicated by a multitude of social, cultural and environmental factors which influence health care decision-making and can otherwise serve to restrict access. We argue that community-engaged research, an orientation which prioritizes collaborative, equitable partnerships and community voice in identifying both problems and solutions, can be a valuable approach to helping address cancer health disparities for Hmong women. Using the Milwaukee-based “Healthy Hmong Women” project as a case example, we detail how the community-engaged approach implemented by the project partners was critical in identifying factors contributing to Hmong cancer disparities and appropriate interventions, as well as the overall acceptance and success of the project. Specifically, we discuss how this approach: (1 promoted community investment and ownership in the project; (2 facilitated the integration of local perspectives and experiences; (3 built capacity to address cancer screening disparities; (4 facilitated the creation of interventions targeting multiple ecological levels; and (5 framed the community as the foundation and driver of positive change.

  10. Investigating Online Destination Images Using a Topic-Based Sentiment Analysis Approach

    Directory of Open Access Journals (Sweden)

    Gang Ren

    2017-09-01

    Full Text Available With the development of Web 2.0, many studies have tried to analyze tourist behavior utilizing user-generated contents. The primary purpose of this study is to propose a topic-based sentiment analysis approach, including a polarity classification and an emotion classification. We use the Latent Dirichlet Allocation model to extract topics from online travel review data and analyze the sentiments and emotions for each topic with our proposed approach. The top frequent words are extracted for each topic from online reviews on Ctrip.com. By comparing the relative importance of each topic, we conclude that many tourists prefer to provide “suggestion” reviews. In particular, we propose a new approach to classify the emotions of online reviews at the topic level utilizing an emotion lexicon, focusing on specific emotions to analyze customer complaints. The results reveal that attraction “management” obtains most complaints. These findings may provide useful insights for the development of attractions and the measurement of online destination image. Our proposed method can be used to analyze reviews from many online platforms and domains.

  11. Investigating Student Perceptions of the Chemistry Laboratory and Their Approaches to Learning in the Laboratory

    Science.gov (United States)

    Berger, Spencer Granett

    This dissertation explores student perceptions of the instructional chemistry laboratory and the approaches students take when learning in the laboratory environment. To measure student perceptions of the chemistry laboratory, a survey instrument was developed. 413 students responded to the survey during the Fall 2011 semester. Students' perception of the usefulness of the laboratory in helping them learn chemistry in high school was related to several factors regarding their experiences in high school chemistry. Students' perception of the usefulness of the laboratory in helping them learn chemistry in college was also measured. Reasons students provided for the usefulness of the laboratory were categorized. To characterize approaches to learning in the laboratory, students were interviewed midway through semester (N=18). The interviews were used to create a framework describing learning approaches that students use in the laboratory environment. Students were categorized into three levels: students who view the laboratory as a requirement, students who believe that the laboratory augments their understanding, and students who view the laboratory as an important part of science. These categories describe the types of strategies students used when conducting experiments. To further explore the relationship between students' perception of the laboratory and their approaches to learning, two case studies are described. These case studies involve interviews in the beginning and end of the semester. In the interviews, students reflect on what they have learned in the laboratory and describe their perceptions of the laboratory environment. In order to encourage students to adopt higher-level approaches to learning in the laboratory, a metacognitive intervention was created. The intervention involved supplementary questions that students would answer while completing laboratory experiments. The questions were designed to encourage students to think critically about the

  12. In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments.

    Science.gov (United States)

    Wu, Chongcong; Nakka, Sravya; Mansouri, Sepahdar; Bengtsson, Torbjörn; Nayeri, Tayeb; Nayeri, Fariba

    2016-09-09

    There is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the host's defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms. The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms. Lymphocytes were separated from whole blood of patients (n = 10) and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system (SPR) and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies. The produced antibodies recognized bacteria with high sensitivity (SPR). The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy. In the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.

  13. Importance of Multifaceted Approaches in Infection Control: A Practical Experience from an Outbreak Investigation.

    Directory of Open Access Journals (Sweden)

    Nina Katharina Stock

    Full Text Available This study presents the results of a multidisciplinary, nosocomial MRSA outbreak investigation in an 8-bed medical intensive care unit (ICU. The identification of seven MRSA positive patients in the beginning of 2014 led to the closure of the ward for several weeks. A multidisciplinary, retrospective investigation was initiated in order to identify the reason and the source for the outbreak, describe MRSA transmission in the department and identify limitations in infection control.The investigation comprised an epidemiological description of MRSA cases from 2012 to 2014 and a characterization of MRSA isolates, including phage-, spa- and PFGE-typing. Additionally, MRSA screening was performed from the hospital staff and the environment. To identify the reason for the outbreak, work-related, psychological and behavioral factors were investigated by impartial audits and staff interviews.Thirty-one MRSA cases were registered during the study period, and 36 isolates were investigated. Molecular typing determined the outbreak strain (phage type 54/812, PFGE type A4, spa type t003 and identified the probable index case. Nasal carriage in one employee and a high environmental contamination with the outbreak strain was documented. Important gaps in nursing procedures and general management were identified. Elevated stress levels and communication problems preceded the outbreak. Compliance with hand hygiene and isolation procedures was evaluated as appropriate.This study demonstrates the complexity of controlling hospital-associated infections. The combined use of different typing methods is beneficial for outbreak investigations. Psychological, behavioral and other work-related factors have an important impact on the spread of nosocomial pathogens. These factors should be addressed and integrated in routine infection control practice.

  14. CFD Approach To Investigate The Flow Characteristics In Bi-Directional Ventilated Disc Brake

    Science.gov (United States)

    Munisamy, Kannan M.; Yusoff, Mohd. Zamri; Shuaib, Norshah Hafeez; Thangaraju, Savithry K.

    2010-06-01

    This paper presents experimental and Computational Fluids Dynamics (CFD) investigations of the flow in ventilated brake discs. Development of an experiment rig with basic measuring devices are detailed out and following a validation study, the possible improvement in the brake cooling can be further analyzed using CFD analysis. The mass flow rate is determined from basic flow measurement technique following that the conventional bi-directional passenger car is simulated using commercial CFD software FLUENT™. The CFD simulation is used to investigate the flow characteristics in between blade flow of the bi-directional ventilated disc brake.

  15. Effects of cryptogamic covers on the global carbon and nitrogen balance as investigated by different approaches

    Science.gov (United States)

    Weber, Bettina; Porada, Philipp; Elbert, Wolfgang; Burrows, Susannah; Caesar, Jennifer; Steinkamp, Jörg; Tamm, Alexandra; Andreae, Meinrat O.; Büdel, Burkhard; Kleidon, Axel; Pöschl, Ulrich

    2014-05-01

    Cryptogamic covers are composed of cyanobacteria, green algae, lichens, bryophytes, fungi and bacteria in varying proportions. As cryptogamic ground covers, comprising biological soil and rock crusts they occur on many terrestrial ground surfaces. Cryptogamic plant covers, containing epiphytic and epiphyllic crusts as well as foliose or fruticose lichens and bryophytes spread over large portions of terrestrial plant surfaces. Photoautotrophic organisms within these crusts sequester atmospheric CO2 and many of them include nitrogen-fixing cyanobacteria, utilizing atmospheric N2 to form ammonium which can be readily used by vascular plants. In a large-scale data analysis approach, we compiled all available data on the physiological properties of cryptogamic covers and developed a model to calculate their annual nitrogen fixation and net primary production. Here, we obtained a total value of 3.9 Pg a-1 for the global net uptake of carbon by cryptogamic covers, which corresponds to approximately 7% of the estimated global net primary production of terrestrial vegetation. Nitrogen assimilation of cryptogamic covers revealed a global estimate of ~49 Tg a-1, accounting for as much as about half the estimated total terrestrial biological nitrogen fixation. In a second approach, we calculated the global carbon uptake by lichens and bryophytes by means of a process-based model. In this model, we used gridded climate data combined with key habitat properties (as e.g. disturbance intervals) to predict the processes which control net carbon uptake, i.e. photosynthesis, respiration, water uptake and evaporation. The model relies on equations frequently used in dynamic vegetation models, which were combined with concepts specific to lichens and bryophytes. As this model only comprises lichens and bryophytes, the predicted terrestrial net uptake of 0.34 to 3.3 Gt a-1 is in accordance with our empirically-derived estimate. Based on this result, we quantified the amount of nitrogen

  16. Cloning Yeast Actin cDNA Leads to an Investigative Approach for the Molecular Biology Laboratory

    Science.gov (United States)

    Black, Michael W.; Tuan, Alice; Jonasson, Erin

    2008-01-01

    The emergence of molecular tools in multiple disciplines has elevated the importance of undergraduate laboratory courses that train students in molecular biology techniques. Although it would also be desirable to provide students with opportunities to apply these techniques in an investigative manner, this is generally not possible in the…

  17. A probabilistic approach to investigate the effect of wave chorology on process-based morphological modelling

    NARCIS (Netherlands)

    Dastgheib, A.; Rajabalinejad, M.R.; Ranasinghe, R.; Roelvink, D.

    2012-01-01

    This paper demonstrates the sensitivity of morphological process-based models to the chronology of input wave conditions. In this research the effect of an emerged offshore breakwater on the morphology of the beach is investigated. A 30 day long morphological simulation with real time history of the

  18. Conventional and Pulse Cathodic Protection of Reinforced Concrete : Electrochemical Approach and Microstructural Investigations

    NARCIS (Netherlands)

    Koleva, D.A.; Hu, J.; Van Breugel, K.; De Wit, J.H.W.; Boshkov, N.

    2006-01-01

    An improved cathodic protection (CP), based on pulse regime was investigated in reinforced concrete, compared to conventional CP. Both regimes used CP current in the range of 2 to15 mA/m2. A voltage-to-current converter maintained the pulse current, the latter adjusted with duty cycle of 12% to 50 %

  19. Investigating Teachers' Understanding of the Salt Dissolution Process: A Multi-Media Approach in Education

    Science.gov (United States)

    Malkoc, Ummuhan

    2017-01-01

    Animations of molecular structure and dynamics are repeatedly applied to support student comprehension in the theoretical ideas of chemistry. However, students' understanding the dynamics of the phenomena is directly related to the understanding of teachers as instructors. Therefore, this study aimed to investigate how the features of three…

  20. Initial investigation of concepts of command and control using an agent based approach

    CSIR Research Space (South Africa)

    Oosthuizen, R

    2010-10-01

    Full Text Available This paper investigates the effect of different Command and Control Concepts (C2) on the outcome of military engagements. The C2 concepts from literature were found to be based on the OODA loop or derivatives thereof. Agent Based Modelling software...

  1. When Child Development Meets Economic Game Theory: An Interdisciplinary Approach to Investigating Social Development

    Science.gov (United States)

    Gummerum, Michaela; Hanoch, Yaniv; Keller, Monika

    2008-01-01

    Game theory has been one of the most prominent theories in the social sciences, influencing diverse academic disciplines such as anthropology, biology, economics, and political science. In recent years, economists have employed game theory to investigate behaviors relating to fairness, reciprocity, and trust. Surprisingly, this research has not…

  2. Investigating the impact of mindfulness meditation training on working memory : A mathematical modeling approach

    NARCIS (Netherlands)

    van Vugt, Marieke K.; Jha, Amishi P.

    We investigated whether mindfulness training (MT) influences information processing in a working memory task with complex visual stimuli. Participants were tested before (T1) and after (T2) participation in an intensive one-month MT retreat, and their performance was compared with that of an age-

  3. DSM-5 AND ICD-11 DEFINITIONS OF POSTTRAUMATIC STRESS DISORDER : INVESTIGATING "NARROW" AND "BROAD" APPROACHES

    NARCIS (Netherlands)

    Stein, Dan J.; McLaughlin, Katie A.; Koenen, Karestan C.; Atwoli, Lukoye; Friedman, Matthew J.; Hill, Eric D.; Maercker, Andreas; Petukhova, Maria; Shahly, Victoria; van Ommeren, Mark; Alonso, Jordi; Borges, Guilherme; de Girolamo, Giovanni; de Jonge, Peter; Demyttenaere, Koen; Florescu, Silvia; Karam, Elie G.; Kawakami, Norito; Matschinger, Herbert; Okoliyski, Michail; Posada-Villa, Jose; Scott, Kate M.; Viana, Maria Carmen; Kessler, Ronald C.

    Background: The development of the Diagnostic and Statistical Manual of Mental Disorders 5th edition (DSM-5) and ICD-11 has led to reconsideration of diagnostic criteria for posttraumatic stress disorder (PTSD). The World Mental Health (WMH) Surveys allow investigation of the implications of the

  4. Single-cell level based approach to investigate bacterial metabolism during batch industrial fermentation

    DEFF Research Database (Denmark)

    Nierychlo, Marta; Larsen, Poul; Eriksen, Niels T.

    culture of Escherichia coli MG1655 was used to investigate glucose and acetate metabolism at single-cell level during different stages of glucose batch fermentation process. Uptake of the substrates was observed and measured in situ by q