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Auswahl der wissenschaftlichen Literatur zum Thema „Permeační studie“
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Zeitschriftenartikel zum Thema "Permeační studie"
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Yeh, Han-I., Jiunn-Tyng Yeh und Tzyh-Chang Hwang. „Modulation of CFTR gating by permeant ions“. Journal of General Physiology 145, Nr. 1 (15.12.2014): 47–60. http://dx.doi.org/10.1085/jgp.201411272.
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Cystic fibrosis transmembrane conductance regulator (CFTR) is unique among ion channels in that after its phosphorylation by protein kinase A (PKA), its ATP-dependent gating violates microscopic reversibility caused by the intimate involvement of ATP hydrolysis in controlling channel closure. Recent studies suggest a gating model featuring an energetic coupling between opening and closing of the gate in CFTR’s transmembrane domains and association and dissociation of its two nucleotide-binding domains (NBDs). We found that permeant ions such as nitrate can increase the open probability (Po) of wild-type (WT) CFTR by increasing the opening rate and decreasing the closing rate. Nearly identical effects were seen with a construct in which activity does not require phosphorylation of the regulatory domain, indicating that nitrate primarily affects ATP-dependent gating steps rather than PKA-dependent phosphorylation. Surprisingly, the effects of nitrate on CFTR gating are remarkably similar to those of VX-770 (N-(2,4-Di-tert-butyl-5-hydroxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide), a potent CFTR potentiator used in clinics. These include effects on single-channel kinetics of WT CFTR, deceleration of the nonhydrolytic closing rate, and potentiation of the Po of the disease-associated mutant G551D. In addition, both VX-770 and nitrate increased the activity of a CFTR construct lacking NBD2 (ΔNBD2), indicating that these gating effects are independent of NBD dimerization. Nonetheless, whereas VX-770 is equally effective when applied from either side of the membrane, nitrate potentiates gating mainly from the cytoplasmic side, implicating a common mechanism for gating modulation mediated through two separate sites of action.
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Piskorowski, Rebecca A., und Richard W. Aldrich. „Relationship between Pore Occupancy and Gating in BK Potassium Channels“. Journal of General Physiology 127, Nr. 5 (24.04.2006): 557–76. http://dx.doi.org/10.1085/jgp.200509482.
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Permeant ions can have significant effects on ion channel conformational changes. To further understand the relationship between ion occupancy and gating conformational changes, we have studied macroscopic and single-channel gating of BK potassium channels with different permeant monovalent cations. While the slopes of the conductance–voltage curve were reduced with respect to potassium for all permeant ions, BK channels required stronger depolarization to open only when thallium was the permeant ion. Thallium also slowed the activation and deactivation kinetics. Both the change in kinetics and the shift in the GV curve were dependent on the thallium passing through the permeation pathway, as well as on the concentration of thallium. There was a decrease in the mean open time and an increase in the number of short flicker closing events with thallium as the permeating ion. Mean closed durations were unaffected. Application of previously established allosteric gating models indicated that thallium specifically alters the opening and closing transition of the channel and does not alter the calcium activation or voltage activation pathways. Addition of a closed flicker state into the allosteric model can account for the effect of thallium on gating. Consideration of the thallium concentration dependence of the gating effects suggests that the flicker state may correspond to the collapsed selectivity filter seen in crystal structures of the KcsA potassium channel under the condition of low permeant ion concentration.
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Tymianski, Michael, Igor Spigelman, Liang Zhang, Peter L. Carlen, Charles H. Tator, Milton P. Charlton und M. Christopher Wallace. „Mechanism of Action and Persistence of Neuroprotection by Cell-Permeant Ca2+ Chelators“. Journal of Cerebral Blood Flow & Metabolism 14, Nr. 6 (November 1994): 911–23. http://dx.doi.org/10.1038/jcbfm.1994.122.
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Cell-permeant Ca2+ chelators such as 1,2-bis-(2-aminophenoxy)ethane- N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM) have been reported to protect neurons in experimental focal cerebral ischemia. However, their in vivo actions are uncertain, and their protective efficacy is proven only in brief cerebral ischemia paradigms. Here we examine their mechanism of action in vitro and duration of efficacy in vivo. Electrophysiological studies were made in CA1 neurons in rat hippocampal slices. When superfused with BAPTA-AM (30–50 μ M), CA1 somatic field potential recordings showed attenuation of the population spike amplitude, and intracellular recordings showed reduced excitatory postsynaptic potentials, indicating inhibition of excitatory synaptic transmission. Also, Ca2+ -dependent accommodation and post-spike-train hyperpolarizations were reduced, indicating Ca2+ chelation hear the internal cell membrane surface. To determine whether Ca2+ chelators reduce the size of cerebral infarction rather than simply delaying its evolution, we studied the effects of BAPTA-AM treatment on infarction size 24 h after permanent middle cerebral artery occlusion. Fischer rats ( n = 8 per group) were pretreated with saline, BAPTA-AM (20 mg/kg), or MK-801 (0.5 mg/kg). Infarction volumes in animals treated with BAPTA-AM were reduced by 50.5% compared with controls ( p = 0.018), whereas animals treated with MK-801 experienced a statistically insignificant infarct volume reduction (26%; p = 0.27). These data show a persistence of neuroprotection by the Ca2+ chelator at 24 h and indicate that it may act by attenuating synaptic transmission and subplasma membrane Ca2+ excess.
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Najib, Omaima N., Stewart B. Kirton, Gary P. Martin, Michelle J. Botha, Al-Sayed Sallam und Darragh Murnane. „Multivariate Analytical Approaches to Identify Key Molecular Properties of Vehicles, Permeants and Membranes That Affect Permeation through Membranes“. Pharmaceutics 12, Nr. 10 (11.10.2020): 958. http://dx.doi.org/10.3390/pharmaceutics12100958.
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There has been considerable recent interest in employing computer models to investigate the relationship between the structure of a molecule and its dermal penetration. Molecular permeation across the epidermis has previously been demonstrated to be determined by a number of physicochemical properties, for example, the lipophilicity, molecular weight and hydrogen bonding ability of the permeant. However little attention has been paid to modeling the combined effects of permeant properties in tandem with the properties of vehicles used to deliver those permeants or to whether data obtained using synthetic membranes can be correlated with those obtained using human epidermis. This work uses Principal Components Analysis (PCA) to demonstrate that, for studies of the diffusion of three model permeants (caffeine, methyl paraben and butyl paraben) through synthetic membranes, it is the properties of the oily vehicle in which they are applied that dominated the rates of permeation and flux. Simple robust and predictive descriptor-based quantitative structure–permeability relationship (QSPR) models have been developed to support these findings by utilizing physicochemical descriptors of the oily vehicles to quantify the differences in flux and permeation of the model compounds. Interestingly, PCA showed that, for the flux of co-applied model permeants through human epidermis, the permeation of the model permeants was better described by a balance between the physicochemical properties of the vehicle and the permeant rather than being dominated solely by the vehicle properties as in the case of synthetic model membranes. The important influence of permeant solubility in the vehicle along with the solvent uptake on overall permeant diffusion into the membrane was substantiated. These results confirm that care must be taken in interpreting permeation data when synthetic membranes are employed as surrogates for human epidermis; they also demonstrate the importance of considering not only the permeant properties but also those of both vehicle and membrane when arriving at any conclusions relating to permeation data.
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Swenson, Erik R., Timothy W. Tewson, Per J. Wistrand, Yvonne Ridderstrale und Chingkuang Tu. „Biochemical, histological, and inhibitor studies of membrane carbonic anhydrase in frog gastric acid secretion“. American Journal of Physiology-Gastrointestinal and Liver Physiology 281, Nr. 1 (01.07.2001): G61—G68. http://dx.doi.org/10.1152/ajpgi.2001.281.1.g61.
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Gastric acid secretion is dependent on carbonic anhydrase (CA). To define the role of membrane-bound CA, we used biochemical, histochemical, and pharmacological approaches in the frog ( Rana pipiens). CA activity and inhibition by membrane-permeant and -impermeant agents were studied in stomach homogenates and microsomal fractions. H+secretion in the histamine-stimulated isolated mucosa was measured before and after mucosal addition of a permeant CA inhibitor (methazolamide) and before and after mucosal or serosal addition of two impermeant CA inhibitors of differing molecular mass: a 3,500-kDa polymer linked to aminobenzolamide and p-fluorobenzyl-aminobenzolamide (molecular mass, 454 kDa). Total CA activity of frog gastric mucosa is 2,280 U/g, of which 10% is due to membrane-bound CA. Membrane-bound CA retains detectable activity below pH 4. Histochemically, there is membrane-associated CA in surface epithelial, oxynticopeptic, and capillary endothelial cells. Methazolamide reduced H+secretion by 100%, whereas the two impermeant inhibitors equally blocked secretion by 40% when applied to the mucosal side and by 55% when applied to the serosal side. The presence of membrane-bound CA in frog oxynticopeptic cells and its relative resistance to acid inactivation and inhibition by impermeant inhibitors demonstrate that it subserves acid secretion at both the apical and basolateral sides.
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Eddins, Donnie, Lisa K. Lyford, Jung Weon Lee, Sanjay A. Desai und Robert L. Rosenberg. „Permeant but not impermeant divalent cations enhance activation of nondesensitizing α7 nicotinic receptors“. American Journal of Physiology-Cell Physiology 282, Nr. 4 (01.04.2002): C796—C804. http://dx.doi.org/10.1152/ajpcell.00453.2001.
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Neuronal α7 nicotinic acetylcholine receptors (nAChRs) are permeable to Ca2+ and other divalent cations. We characterized the modulation of the pharmacological properties of nondesensitizing mutant (L247T and S240T/L247T) α7 nAChRs by permeant (Ca2+, Ba2+, and Sr2+) and impermeant (Cd2+ and Zn2+) divalent cations. α7 receptors were expressed in Xenopus oocytes and studied with two-electrode voltage clamp. Extracellular permeant divalent cations increased the potency and maximal efficacy of ACh, whereas impermeant divalent cations decreased potency and maximal efficacy. The antagonist dihydro-β-erythroidine (DHβE) was a strong partial agonist of L247T and S240T/L247T α7 receptors in the presence of divalent cations but was a weak partial agonist in the presence of impermeant divalent cations. Mutation of the “intermediate ring” glutamates (E237A) in L247T α7 nAChRs eliminated Ca2+conductance but did not alter the Ca2+-dependent increase in ACh potency, suggesting that site(s) required for modulation are on the extracellular side of the intermediate ring. The difference between permeant and impermeant divalent cations suggests that sites within the pore are important for modulation by divalent cations.
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Thompson, Jill, und Ted Begenisich. „Selectivity filter gating in large-conductance Ca2+-activated K+ channels“. Journal of General Physiology 139, Nr. 3 (27.02.2012): 235–44. http://dx.doi.org/10.1085/jgp.201110748.
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Membrane voltage controls the passage of ions through voltage-gated K (Kv) channels, and many studies have demonstrated that this is accomplished by a physical gate located at the cytoplasmic end of the pore. Critical to this determination were the findings that quaternary ammonium ions and certain peptides have access to their internal pore-blocking sites only when the channel gates are open, and that large blocking ions interfere with channel closing. Although an intracellular location for the physical gate of Kv channels is well established, it is not clear if such a cytoplasmic gate exists in all K+ channels. Some studies on large-conductance, voltage- and Ca2+-activated K+ (BK) channels suggest a cytoplasmic location for the gate, but other findings question this conclusion and, instead, support the concept that BK channels are gated by the pore selectivity filter. If the BK channel is gated by the selectivity filter, the interactions between the blocking ions and channel gating should be influenced by the permeant ion. Thus, we tested tetrabutyl ammonium (TBA) and the Shaker “ball” peptide (BP) on BK channels with either K+ or Rb+ as the permeant ion. When tested in K+ solutions, both TBA and the BP acted as open-channel blockers of BK channels, and the BP interfered with channel closing. In contrast, when Rb+ replaced K+ as the permeant ion, TBA and the BP blocked both closed and open BK channels, and the BP no longer interfered with channel closing. We also tested the cytoplasmically gated Shaker K channels and found the opposite behavior: the interactions of TBA and the BP with these Kv channels were independent of the permeant ion. Our results add significantly to the evidence against a cytoplasmic gate in BK channels and represent a positive test for selectivity filter gating.
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Yoshikawa, Masakazu, Takeshi Matsuura und David Cooney. „Studies on the state of permeant in the membrane and its effect on pervaporation phenomena“. Journal of Applied Polymer Science 42, Nr. 5 (05.03.1991): 1417–21. http://dx.doi.org/10.1002/app.1991.070420526.
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Otto, Daniel, Johann Combrinck, Anja Otto, Louwrens Tiedt und Melgardt de Villiers. „Dissipative Particle Dynamics Investigation of the Transport of Salicylic Acid through a Simulated In Vitro Skin Permeation Model“. Pharmaceuticals 11, Nr. 4 (05.12.2018): 134. http://dx.doi.org/10.3390/ph11040134.
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Permeation models are often used to determine diffusion properties of a drug through a membrane as it is released from a delivery system. In order to circumvent problematic in vivo studies, diffusion studies can be performed in vitro, using (semi-)synthetic membranes. In this study salicylic acid permeation was studied, employing a nitrocellulose membrane. Both saturated and unsaturated salicylic acid solutions were studied. Additionally, the transport of salicylic acid through the nitrocellulose membrane was simulated by computational modelling. Experimental observations could be explained by the transport mechanism that was revealed by dissipative particle dynamics (DPD) simulations. The DPD model was developed with the aid of atomistic scale molecular dynamics (AA-MD). The choice of a suitable model membrane can therefore, be predicted by AA-MD and DPD simulations. Additionally, the difference in the magnitude of release from saturated and unsaturated salicylic acid and solutions could also be observed with DPD. Moreover, computational studies can reveal hidden variables such as membrane-permeant interaction that cannot be measured experimentally. A recommendation is made for the development of future model permeation membranes is to incorporate computational modelling to aid the choice of model.
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Shapiro, M. S., und T. E. DeCoursey. „Permeant ion effects on the gating kinetics of the type L potassium channel in mouse lymphocytes.“ Journal of General Physiology 97, Nr. 6 (01.06.1991): 1251–78. http://dx.doi.org/10.1085/jgp.97.6.1251.
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Permeant ion species was found to profoundly affect the gating kinetics of type l K+ currents in mouse T lymphocytes studied with the whole-cell or on-cell patch gigaohm-seal techniques. Replacing external K+ with Rb+ (as the sole monovalent cation, at 160 mM) shifted the peak conductance voltage (g-V) relation by approximately 20 mV to more negative potentials, while NH4+ shifted the g-V curve by 15 mV to more positive potentials. Deactivation (the tail current time constant, tau tail) was slowed by an average of 14-fold at -70 mV in external Rb+, by approximately 8-fold in Cs+, and by a factor of two to three in NH4+. Changing the external K+ concentration, [K+]o, from 4.5 to 160 mM or [Rb+]o from 10 to 160 mM had no effect on tau tail. With all the internal K+ replaced by Rb+ or Cs+ and either isotonic Rb+ or K+ in the bath, tau tail was indistinguishable from that with K+ in the cell. With the exception of NH4+, activation time constants were insensitive to permeant ion species. These results indicate that external permeant ions have stronger effects than internal permeant ions, suggesting an external modulatory site that influences K+ channel gating. However, in bi-ionic experiments with reduced external permeant ion concentrations, tau tail was sensitive to the direction of current flow, indicating that the modulatory site is either within the permeation pathway or in the outer vestibule of the channel. The latter interpretation implies that outward current through an open type l K+ channel significantly alters local ion concentrations at the modulatory site in the outer vestibule, and consequently at the mouth of the channel. Experiments with mixtures of K+ and Rb+ in the external solution reveal that deactivation kinetics are minimally affected by addition of Rb+ until the Rb+ mole fraction approaches unity. This relationship between mole fraction and tau tail, together with the concentration independence of tau tail, was hard to reconcile with simple models in which occupancy of a site within the permeation pathway prevents channel closing, but is consistent with a model in which a permeant ion binding site in the outer vestibule modulates gating depending on the species of ion occupying the site. A description of the ionic selectivity of the type l K+ channel is presented in the companion paper (Shapiro and DeCoursey, 1991b).
Dissertationen zum Thema "Permeační studie"
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Almansour, Khaled. „Mechanistic studies of a cell-permeant peptide designed to enhance myosin light chain phosphorylation“. Thesis, University of Bath, 2017. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.760954.
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Oral delivery of therapeutic peptides has been a continuous target for the pharmaceutical industry, as most of these drugs are currently administered by parenteral routes. However, a major challenge limiting the success of oral delivery of these drugs is their poor permeability across the intestinal epithelial barrier. Extensive research efforts have investigated different strategies to overcome the epithelial barrier and enhance the oral bioavailability of therapeutic peptides. One of the most widely used strategies is the application of permeability enhancer (PE) agents that are co-administered with peptide drugs to facilitate their permeability across the intestinal epithelial barrier. The safety of most of the available PE agents, however, has always been questioned, because most PE agents act non-specifically in altering intestinal epithelial permeability which in many cases has been associated with epithelial damage. The work presented here investigates a novel strategy to overcome the intestinal epithelial barrier challenge and enhance the oral bioavailability of therapeutic peptides. This is by manipulating an endogenous mechanism that is used by the intestinal epithelial cells to dynamically regulate the permeability across the tight junction (TJ) structures by increasing myosin light chain phosphorylation at serine 19 (MLC-pS19), which is regulated by MLC kinase (MLCK) and MLC phosphatase (MLCP). A small membrane-permeant peptide inhibitor for MLCP, called PIP 640 peptide, was rationally designed to selectively alter MLCP activity in a manner that increases MLC-pS19 to transiently enhance TJ permeability for therapeutic peptides. The PIP 640 peptide was designed to be relatively stable in the intestinal lumen, as it is intended to be co-administered orally with therapeutic peptides. It was initially examined for enhanced TJ permeability of fluorescent dextran and for toxicity induction in vitro. Accordingly, efforts were devoted to explore potential modifications of the peptide sequence that might optimize the PIP 640 peptide function. Moreover, studies were performed to examine the biochemical changes of TJ protein structures associated with the permeability enhancement function of the PIP 640 peptide. Finally, we investigated different aspects of the PIP 640 peptide permeability enhancement function in vivo. An overall outcome of these studies was that the PIP 640 peptide can enhance TJ permeability in vitro and in vivo without causing apparent damage to the epithelial barrier. This outcome suggest that the PIP 640 peptide has the potential to be used as a PE for therapeutic peptides.
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Owens, Sian-Eleri. „Studies of myosin light chain-dependent modulation of tight junction function through the actions of membrane-permeant peptides“. Thesis, Cardiff University, 2007. http://orca.cf.ac.uk/55667/.
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Myosin light chain phosphorylation plays a central role in the regulation of paracellular permeability. The main objective of this study was to design and synthesise membrane-permeant peptide inhibitors of myosin light chain kinase and phosphatase that could potentially decrease and increase paracellular permeability, respectively. Elevated myosin light chain kinase activity, as observed in a variety of inflammatory disease, phosphorylates myosin light chain, to increase paracellular permeability. Initial studies showed a peptide inhibitor of myosin light chain kinase termed PIK could rectify increased paracellular permeability in two different cell-based models of inflammation. PIK was also, however, shown to be too labile for use in vivo. From a series of PIK analogues, two candidates prepared using D-amino acids were identified that showed sufficient stability, membrane-permeant properties and retention of specific function of PIK for future in vivo studies. Since the ratio of myosin light chain kinase to phosphatase activity regulates the degree of myosin light chain phosphorylation, it was hypothesised that inhibition of myosin light chain phosphatase would increase paracellular permeability. A membrane-permeant peptide designed to inhibit myosin light chain phosphatase activity termed PIP was identified in a screen of potential candidates through its capability to significantly decrease transepithelial electrical resistance in a polarized human intestinal epithelial cell system in vitro without any apparent cytotoxicity. Further studies will be required to determine the extent to which PIP increases paracellular permeability. Using these novel membrane-permeant peptide inhibitors, studies were performed using polarized monolayers of human intestinal epithelial cells to assess the action of previously identified regulators of paracellular permeability in vitro. Previous studies demonstrated that the non-specific myosin light chain kinase inhibitor ML-7 prevented the absorption enhancing properties of sodium caprate. Treatment with PIK and sodium caprate simultaneously resulted in significant increases in the permeability of inert fluorescent probes while pre- and post-incubation with PIK inhibited sodium caprate effects. These surprising findings suggest a potential application for combination treatment with sodium caprate and PIK to increase paracellular permeability of poorly absorbed drugs.
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Nedbálková, Iva. „Permeační in vitro studie stability transkarbamu 12 v oleokrémech“. Master's thesis, 2008. http://www.nusl.cz/ntk/nusl-292548.
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The work extends the original method of biological titration with the concrete assessment of the stability of an active form of T 12 in two creams of functionally usable composition. The work compiles basic information on the skin barrier, chemically modulated penetration of drugs through the skin, especially transkarbam 12 (T 12) as an accelerator. Its influence on the permeation characteristics of caffeine as a marker is a basis of the experiment itself. The principal experimental results have been obtained by measuring of in vitro (pig skin ear samples) fluxes of caffeine dispersed always in 1% concentration in hydrophobic creams containing 0.2% and 0.4% of T 12. The average results of the pertinent fluxes were for the 0.2% cream J = 6,4 ± 2,0 in one week after manufacturing, and J = 7,2 ± 2,3 in 210 days after manufacturing. Analogical values for the 0.4% lotion were J = 6,7 ± 1, in one week after manufacturing, and J = 5,5 ± 1,2 after 165 days after manufacturing. The evaluation of these results by non-pair t-tests with the level of reliability of 0.95% has proved that the permeation characteristics of both creams (with 0.2% and 0.4% of T 12) in the scrutinized period (210 days or 165 days, respectively). It is thus possible to treat them as equal from the permeation point of view, and the...
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Šimek, Matěj. „In vitro modely kožní bariéry“. Master's thesis, 2021. http://www.nusl.cz/ntk/nusl-445839.
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Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Technology Candidate: Matěj Šimek Consultant: doc. Dr. rer. nat. Mgr. Jarmila Zbytovská Title of thesis: In vitro models of skin barrier The aim of this work is to summarize information about various types of skin models which are used for testing of permeability, toxicity, irritability and other aspects of drugs, through professional, verified and reviewed literature. These characteristics are necessary to know in order to grant optimal safety, effectiveness nad quality of transdermally administered drugs. Transdermal administration of drugs has got lots of benefits in contrast with classic peroral administration. An administration of drugs through this way is quite simple and it can be interrupted quite easily. We can also easily change a place of administration in which a drug is released and the risk of overdosing is very low. Furthermore, transdermal administration makes possible to maintain constant plasmatic concentration of drug in a blood stream and also to prolong the duration of effect of drugs with small halftime thanks to constant releasing of drug. And primarily, transdermally administered drug normally avoids the "first-pass" effect of liver, so the dose of drug can be lowered. The risk of drug...
Bücher zum Thema "Permeační studie"
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Krapova, Iliyana, Svetlana Nistratova und Luisa Ruvoletto. Studi di linguistica slava. Venice: Edizioni Ca' Foscari, 2019. http://dx.doi.org/10.30687/978-88-6969-368-7.
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I contributi raccolti nel presente volume delineano lo stato dell’arte delle ricerche di linguistica slava svolte recentemente nell’ambito della slavistica italiana. I saggi sono dedicati a temi di morfologia, sintassi, semantica, lessicologia, pragmatica, sociolinguistica e didattica delle lingue slave, in ottica contrastiva, sincronica o diacronica, secondo quadri teorici e approcci metodologici di scuole e tradizioni diverse. La grande varietà dei temi trattati dagli autori, non solo italiani, è la più viva testimonianza della vivacità e della ricchezza che oggi permeano lo studio delle lingue slave in Italia e non solo.