Effects of Experimental Setup on the Apparent Concentration Dependency of Active Efflux Transport in in Vitro Cell Permeation Experiments

The immunofluorescence technique was used to detect the presence and distribution of actin, alpha-actinin, tubulin and 10 nm filament protein in early mouse embryos. Actin and alpha-actinin stainings showed a distinct concentration to a peripheral layer in the cleavage-stage blastomeres and in trophectoderm cells. Dots of fluorescence appeared in this cortical staining pattern. The distribution of tubulin staining in the blastomere cytoplasm was relatively even with apparent concentration at the perinuclear region and frequently at wide intercellular contact areas. 10 nm filament protein was distributed evenly in the blastomere cytoplasm without cortical concentration of the label. At the blastocyst stage, the trophectoderm cells in blastocyst outgrowths in vitro developed well organized cytoskeletons including both microfilament, microtubule and 10 nm filament elements. Comparable structures were not observed in blastocysts in vivo, or in late hatched blastocysts cultured in suspension. The morphogenetic significance of the observations is discussed.

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Water Jet Applicator for Interface Tissue Removal in Minimally Invasive Hip Refixation: Testing the Principle and Design of Prototype

Journal of Medical Devices ◽

10.1115/1.4043293 ◽

2019 ◽

Vol 13 (2) ◽

Author(s):

Gert Kraaij ◽

Arjo J. Loeve ◽

Jenny Dankelman ◽

Rob G. H. H. Nelissen ◽

Edward R. Valstar

Keyword(s):

Theoretical Analysis ◽

Minimally Invasive ◽

Water Pressure ◽

Minimal Invasive ◽

Outer Diameter ◽

Experimental Setup ◽

Technical Requirements ◽

Tissue Removal ◽

Further Development

Mechanical loosening of implants is in the majority accompanied with a periprosthetic interface membrane, which has to be removed during revision surgery. The same is true if a minimal invasive (percutaneous) refixation of a loose implant is done. We describe the requirements for a waterjet applicator for interface tissue removal for this percutaneous hip refixation technique. The technical requirements were either obtained from a literature review, a theoretical analysis, or by experimental setup. Based on the requirements, a waterjet applicator is designed which is basically a flexible tube (outer diameter 3 mm) with two channels. One channel for the water supply (diameter 0.9 mm) and one for suction to evacuate water and morcellated interface tissue from the periprosthetic cavity. The applicator has a rigid tip (length 6 mm), which directs the water flow to create two waterjets (diameter 0.2 mm), both focused into the suction channel. The functionality of this new applicator is demonstrated by testing a prototype of the applicator tip in an in vitro experimental setup. This testing has shown that the designed applicator for interface tissue removal will eliminate the risk of water pressure buildup; the ejected water was immediately evacuated from the periprosthetic cavity. Blocking of the suction opening was prevented because the jets cut through interface tissue that gets in front of the suction channel. Although further development of the water applicator is necessary, the presented design of the applicator is suitable for interface tissue removal in a minimally invasive hip refixation procedure.

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In-vitro cell treatment with focused shockwaves—influence of the experimental setup on the sound field and biological reaction

Journal of Therapeutic Ultrasound ◽

10.1186/s40349-016-0053-z ◽

2016 ◽

Vol 4 (1) ◽

Cited By ~ 10

Author(s):

Kristin Dietz-Laursonn ◽

Rainer Beckmann ◽

Siegfried Ginter ◽

Klaus Radermacher ◽

Matías de la Fuente

Keyword(s):

Sound Field ◽

Experimental Setup ◽

Biological Reaction

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One Dimensional Experimental Setup to Study the Heating of Nanoparticle Laden Systems

ASME 2010 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2010-19676 ◽

2010 ◽

Cited By ~ 3

Author(s):

Zhenpeng Qin ◽

John C. Bischof

Keyword(s):

Small Volume ◽

Laser Energy ◽

Special Kind ◽

Gold Nanoshells ◽

Experimental Setup ◽

Heat Absorption ◽

One Dimensional ◽

Lumped Models

Intensive efforts have been put into the use of gold nanoparticles (GNPs) for the enhancement of hyperthermia using laser in recent years since the groundbreaking work of Hirsh et al.(1) using gold nanoshells (GNS). Both in vitro (2), and in vivo (3) studies show promising results. For example, GNS, a special kind of GNP, are being manufactured and are in clinical trials (Nanospectra Bioscience, Inc). While the data is compelling, unfortunately the fundamentals of GNP heating are not entirely understood. For example, there are large discrepancies in the experimentally measured photothermal efficiency of GNPs (4, 5). Furthermore, lumped models of GNP heating in solution, by using small volume of GNP solution (4, 5), or stirring the solution (6), neglecting the variation of heat absorption throughout a system require improvement. In reality, the GNPs will attenuate the laser beam as it passes through the GNP host medium. GNPs at different locations will absorb different amount of laser energy and hence have different heat generation.

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In Vitro Activities of Garenoxacin (BMS-284756) against Streptococcus pneumoniae, Viridans Group Streptococci, and Enterococcus faecalis Compared to Those of Six Other Quinolones

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.47.11.3542-3547.2003 ◽

2003 ◽

Vol 47 (11) ◽

pp. 3542-3547 ◽

Cited By ~ 19

Author(s):

Patrick Grohs ◽

Serge Houssaye ◽

Agnès Aubert ◽

Laurent Gutmann ◽

Emmanuelle Varon

Keyword(s):

Streptococcus Pneumoniae ◽

Enterococcus Faecalis ◽

Bactericidal Effect ◽

Clinical Isolates ◽

Active Efflux ◽

Resistant Strains ◽

Different Strains ◽

Viridans Group Streptococci ◽

Derivatives Of

ABSTRACT The activity of garenoxacin, a new quinolone, was determined in comparison with other quinolones against different strains of S. pneumoniae, viridans group streptococci (VGS), and Enterococcus faecalis. Strains were quinolone-susceptible clinical isolates and quinolone-resistant strains with defined mechanisms of resistance obtained from either clinical isolates or derivatives of S. pneumoniae R6. Clinical quinolone-susceptible strains of S. pneumoniae, VGS and E. faecalis showed garenoxacin MICs within a range of 0.03 μg/ml to 0.25 μg/ml. Garenoxacin MICs increased two- to eightfold when one mutation was present in the ParC quinolone resistance-determining region (QRDR), fourfold when one mutation was present in the GyrA QRDR (S. pneumoniae), 8- to 64-fold when two or three mutations were associated in ParC and GyrA QRDR, and 2,048-fold when two mutations were present in both the GyrA and ParC QRDRs (Streptococcus pneumoniae). Increased active efflux had a moderate effect on garenoxacin MICs for S. pneumoniae and VGS. Against S. pneumoniae, garenoxacin behaved like moxifloxacin and sparfloxacin, being more affected by a single gyrA mutation than by a single parC mutation. Although garenoxacin was generally two- to fourfold more active than moxifloxacin against the different wild-type or mutant strains of S. pneumoniae, VGS, and E. faecalis, it was two- to fourfold less active than gemifloxacin. At four times the respective MIC for each strain, the bactericidal effect of garenoxacin, observed at 6 h for S. pneumoniae and at 24 h for S. oralis and E. faecalis, was not influenced by the presence of mutation either in the ParC or in both the ParC and GyrA QRDRs.

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Physiologically Based Pharmacokinetic Modeling of Rosuvastatin to Predict Transporter-Mediated Drug-Drug Interactions

Pharmaceutical Research ◽

10.1007/s11095-021-03109-6 ◽

2021 ◽

Author(s):

Nina Hanke ◽

José David Gómez-Mantilla ◽

Naoki Ishiguro ◽

Peter Stopfer ◽

Valerie Nock

Keyword(s):

Drug Interactions ◽

Pbpk Model ◽

Open Systems ◽

Whole Body ◽

Good Prediction ◽

Active Efflux ◽

Physiologically Based Pharmacokinetic ◽

Positron Emission ◽

Physiologically Based

Abstract Purpose To build a physiologically based pharmacokinetic (PBPK) model of the clinical OATP1B1/OATP1B3/BCRP victim drug rosuvastatin for the investigation and prediction of its transporter-mediated drug-drug interactions (DDIs). Methods The Rosuvastatin model was developed using the open-source PBPK software PK-Sim®, following a middle-out approach. 42 clinical studies (dosing range 0.002–80.0 mg), providing rosuvastatin plasma, urine and feces data, positron emission tomography (PET) measurements of tissue concentrations and 7 different rosuvastatin DDI studies with rifampicin, gemfibrozil and probenecid as the perpetrator drugs, were included to build and qualify the model. Results The carefully developed and thoroughly evaluated model adequately describes the analyzed clinical data, including blood, liver, feces and urine measurements. The processes implemented to describe the rosuvastatin pharmacokinetics and DDIs are active uptake by OATP2B1, OATP1B1/OATP1B3 and OAT3, active efflux by BCRP and Pgp, metabolism by CYP2C9 and passive glomerular filtration. The available clinical rifampicin, gemfibrozil and probenecid DDI studies were modeled using in vitro inhibition constants without adjustments. The good prediction of DDIs was demonstrated by simulated rosuvastatin plasma profiles, DDI AUClast ratios (AUClast during DDI/AUClast without co-administration) and DDI Cmax ratios (Cmax during DDI/Cmax without co-administration), with all simulated DDI ratios within 1.6-fold of the observed values. Conclusions A whole-body PBPK model of rosuvastatin was built and qualified for the prediction of rosuvastatin pharmacokinetics and transporter-mediated DDIs. The model is freely available in the Open Systems Pharmacology model repository, to support future investigations of rosuvastatin pharmacokinetics, rosuvastatin therapy and DDI studies during model-informed drug discovery and development (MID3).

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Torque Loss After Miniscrew Placement: An In-Vitro Study Followed by a Clinical Trial

The Open Dentistry Journal ◽

10.2174/1874210601610010251 ◽

2016 ◽

Vol 10 (1) ◽

pp. 251-260 ◽

Cited By ~ 2

Author(s):

Marco Migliorati ◽

Sara Drago ◽

Fabrizio Barberis ◽

Irene Schiavetti ◽

Domenico Dalessandri ◽

...

Keyword(s):

In Vitro Study ◽

Insertion Time ◽

Controlled Environment ◽

Insertion Torque ◽

Experimental Setup ◽

Percentage Difference ◽

Torque Loss ◽

Torque Values

To evaluate torque loss a week after insertion, both in an in vivo and an in vitro experimental setup were designed. In the in vivo setup a total of 29 miniscrews were placed in 20 patients who underwent orthodontic treatment. Maximum insertion torque (MIT) was evaluated at insertion time (T1). A week later, insertion torque was measured again by applying a quarter turn (T2); no load was applied on the screw during the first week. In the in vitro setup a total of 20 miniscrews were placed in pig rib bone samples. MIT was evaluated at insertion time (T1). Bone samples were kept in saline solution and controlled environment for a week during which the solution was refreshed every day. Afterwards, torque was measured again by applying a quarter turn (T2). The comparison of MIT over time was done calculating the percentage difference of the torque values between pre- and post-treatment and using the parametric two independent samples t-test or the non-parametric Mann–Whitney test. After a week unloaded miniscrews showed a mean loss of rotational torque of 36.3% and 40.9% in in vitro and in in vivo conditions, respectively. No statistical differences were found between the two different setups. Torque loss was observed after the first week in both study models; in vitro experimental setup provided a reliable study model for studying torque variation during the first week after insertion.

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Catheter tip distensibility substantially influences the aspiration force of thrombectomy devices

Journal of NeuroInterventional Surgery ◽

10.1136/neurintsurg-2021-017487 ◽

2021 ◽

pp. neurintsurg-2021-017487

Author(s):

Jiahui Li ◽

Oscar Castaño ◽

Alejandro Tomasello ◽

Marta de Dios Lascuevas ◽

Pere Canals ◽

...

Keyword(s):

Relative Difference ◽

Experimental Setup ◽

Section Area ◽

Wide Range ◽

Device Efficiency ◽

First Pass ◽

Catheter Tip ◽

3D Printed ◽

Inner Diameter

BackgroundA direct aspiration first pass thrombectomy (ADAPT) is a fast-growing technique for which a broad catalog of catheters that provide a wide range of aspiration forces can be used. We aimed to characterize different catheters' aspiration performance on stiff clots in an in vitro vascular model. We hypothesized that labeled catheter inner diameter (labeled-ID) is not the only parameter that affects the aspiration force (asp-F) and that thrombus–catheter tip interaction and distensibility also play a major role.MethodsWe designed an experimental setup consisting of a 3D-printed carotid artery immersed in a water deposit. We measured asp-F and distensibility of catheter tips when performing ADAPT on a stiff clot analog larger than catheter labeled-ID. Correlations between asp-F, catheter ID, and tip distensibility were statistically assessed.ResultsExperimental asp-F and catheter labeled-ID were correlated (r=0.9601; P<0.01). The relative difference between experimental and theoretical asp-F (obtained by the product of the tip’s section area by the vacuum pressure) correlated with tip’s distensibility (r=0.9050; P<0.01), evidencing that ADAPT performance is highly influenced by catheter tip shape-adaptability to the clot and that the effective ID (eff-ID) may differ from the labeled-ID specified by manufacturers. Eff-ID showed the highest correlation with experimental asp-F (r=0.9944; P<0.01), confirming that eff-ID rather than labeled-ID should be considered to better estimate the device efficiency.ConclusionsCatheter tip distensibility can induce a significant impact on ADAPT performance when retrieving a stiff clot larger than the device ID. Our findings might contribute to optimizing thrombectomy strategies and the design of novel aspiration catheters.

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Kinetics of active efflux via choroid plexus of beta-lactam antibiotics from the CSF into the circulation

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1994.266.2.r392 ◽

1994 ◽

Vol 266 (2) ◽

pp. R392-R399 ◽

Cited By ~ 2

Author(s):

M. Ogawa ◽

H. Suzuki ◽

Y. Sawada ◽

M. Hanano ◽

Y. Sugiyama

Keyword(s):

Choroid Plexus ◽

Maximum Velocity ◽

Organic Anions ◽

Dependent Manner ◽

Beta Lactam ◽

Active Efflux ◽

Beta Lactam Antibiotics ◽

Elimination Clearance

To examine the role of the choroid plexus in eliminating organic anions from the cerebrospinal fluid (CSF), a kinetic study was performed both in in vivo and in vitro experiments using [3H]benzylpenicillin (PCG) as a model compound. In vivo, after intracerebroventricular administration, [3H]PCG was eliminated from the CSF much more rapidly than [14C]mannitol. Analysis of the elimination clearance from the CSF revealed that 12 and 24% of the disappearance of [3H]PCG can be accounted for by convective loss at a rate equivalent to CSF turnover, and by diffusional loss across the ependymal surface into the brain extracellular space, respectively. Approximately two-thirds of [3H]PCG elimination was due to a saturable process [Michaelis constant (Km) = 43.0 +/- 17.8 microM, maximum velocity (Vmax) = 619 +/- 286 pmol.min-1 x rat-1]. These kinetic parameters obtained in vivo were comparable to those determined previously in vitro, i.e., [3H]PCG was accumulated by the isolated rat choroid plexus via an active transport mechanism (Km = 58 microM, Vmax = 504 pmol.min-1 x rat-1; H. Suzuki, Y. Sawada, Y. Sugiyama, T. Iga, and H. Hanano, J. Pharmacol. Exp. Ther. 242: 660-665, 1987). Furthermore, other organic anions (probenecid, ampicillin, cefodizime, cefotaxime, and ceftriaxone) reduced the transport of [3H]PCG in a dose-dependent manner both in vivo and in vitro. A good correlation was observed between the log inhibition constant (Ki) values obtained for these ligands in vivo and in vitro (r = 0.94, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

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