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Membrane Permeability (membrane + permeability)
Kinds of Membrane Permeability Selected AbstractsEnhancing Membrane Permeability by Fatty Acylation of Oligoarginine PeptidesCHEMBIOCHEM, Issue 8 2004Wellington Pham Dr. A systematic study of fatty acylated polyarginines as drug-delivery modules is described. A remarkable uptake was observed when a myristate moiety was incorporated with a 7-mer polyarginine (see scheme). Microscopic studies suggest that the fatty acylated peptide is internalized into the cytoplasm, not the nucleus, thus providing an alternative method for drug delivery. [source] Wheat Cellular Membrane Thermotolerance Under Heat StressJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 2 2010A. S. Dias Abstract Four genotypes of Triticum aestivum L. and Triticum turgidum subsp. durum chosen according to their genetic background diversity were subjected to heat stress after anthesis. Membrane permeability, lipid peroxidation and fatty acids (C14:0, C16:0, C16:1c, C16:1t, C18:0, C18:1, C18:2 and C18:3) were quantified. The estimation of the quantum yield of non-cyclic photosynthetic electron transport was used as well as a test system to further evaluate the implications on thylakoid functioning. It was found differences within bread and durum wheat species concerning the capability to cope with high temperatures at the stage of grain filling. The genotype Sever showed high thermal sensitivity concerning membrane lipid peroxidation and membrane permeability, as evaluated by the increased production of ethylene and MDA, as well as by the impact on TFA (at the middle term of grain filling). In the durum wheat genotypes, differences were also found, with TE 9306 displaying high membrane stability, with no increases on membrane permeability, MDA and ethylene content. In this way, the observed changes on TFA in this genotype might have constituted a mechanism to allow qualitative lipid changes, reflected in lower unsaturation level of membrane FAs which is a positive trait under high temperatures. [source] Membrane permeability and antimicrobial kinetics of cecropin P1 against Escherichia coli,JOURNAL OF PEPTIDE SCIENCE, Issue 6 2009Steven Arcidiacono Abstract The interaction of cecropin P1 (CP1) with Escherichiacoli was investigated to gain insight into the time-dependent antimicrobial action. Biophysical characterizations of CP1 with whole bacterial cells were performed using both fluorescent and colorimetric assays to investigate the role of membrane permeability and lipopolysaccharide (LPS) binding in lytic behavior. The kinetics of CP1 growth inhibition assays indicated a minimal inhibitory concentration (MIC) of 3 µM. Bactericidal kinetics at the MIC indicated rapid killing of E.coli (<30 min). Membrane permeability studies illustrated permeation as a time-dependent event. Maximum permeability at the MIC occurred within 30 min, which correlates to the bactericidal action. Further investigation showed that the immediate permeabilizing action of CP1 is concentration-dependent, which correlates to the concentration-dependent nature of the inhibition assays. At the MIC and above, the immediate permeability was significant enough that the cells could not recover and exhibit growth. Below the MIC, immediate permeability was evident, but the level was insufficient to inhibit growth. Dansyl polymyxin B displacement studies showed LPS binding is essentially the same at all concentrations investigated. However, it does appear that only the immediate interaction is important, because binding continued to increase over time beyond cell viability. Our studies correlated CP1 bactericidal kinetics to membrane permeability suggesting CP1 concentration-dependent killing is driven by the extent of the immediate permeabilizing action of the peptide. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd. [source] The novel herbicide oxaziclomefone inhibits cell expansion in maize cell cultures without affecting turgor pressure or wall acidificationNEW PHYTOLOGIST, Issue 2 2005Nichola O'Looney Summary ,,Oxaziclomefone [OAC; IUPAC name 3-(1-(3,5-dichlorophenyl)-1-methylethyl)-3,4-dihydro-6-methyl-5-phenyl-2H -1,3-oxazin-4-one] is a new herbicide that inhibits cell expansion in grass roots. Its effects on cell cultures and mode of action were unknown. In principle, cell expansion could be inhibited by a decrease in either turgor pressure or wall extensibility. ,,Cell expansion was estimated as settled cell volume; cell division was estimated by cell counting. Membrane permeability to water was measured by a novel method involving simultaneous assay of the efflux of 3H2O and [14C]mannitol from a ,bed' of cultured cells. Osmotic potential was measured by depression of freezing point. ,,OAC inhibited cell expansion in cultures of maize (Zea mays), spinach (Spinacia oleracea) and rose (Rosa sp.), with an ID50 of 5, 30 and 250 nm, respectively. In maize cultures, OAC did not affect cell division for the first 40 h. It did not affect the osmotic potential of cell sap or culture medium, nor did it impede water transport across cell membranes. It did not affect cells' ability to acidify the apoplast (medium), which may be necessary for ,acid growth'. ,,As OAC did not diminish turgor pressure, its ability to inhibit cell expansion must depend on changes in wall extensibility. It could be a valuable tool for studies on cell expansion. [source] Optimization of mass transfer for toxin removal and immunoprotection of hepatocytes in a bioartificial liverBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2009Geir I. Nedredal Abstract This study was designed to determine optimal operating conditions of a bioartificial liver (BAL) based on mass transfer of representative hepatotoxins and mediators of immune damage. A microprocessor-controlled BAL was used to study mass transfer between patient and cell compartments separated by a hollow fiber membrane. Membrane permeability (70, 150, or 400,kDa molecular weight cut-off,MWCO), membrane convection (high: 50,mL/min; medium: 25,mL/min; low: 10,mL/min; diffusion: 0,mL/min), and albumin concentration in the cell compartment (0.5 or 5,g%) were considered for a total of 24 test conditions. Initially, the patient compartment contained pig plasma supplemented with ammonia (0.017,kDa), unconjugated bilirubin (0.585,kDa), conjugated bilirubin (0.760,kDa), TNF-, (17,kDa), pig albumin (67,kDa), pig IgG (147,kDa), and pig IgM (900,kDa). Mass transfer of each substance was determined by its rate of appearance in the cell compartment. Membrane fouling was assessed by dextran polymer technique. Of the three tested variables (membrane pore size, convection, and albumin concentration), membrane permeability had the greatest impact on mass transfer (P,<,0.001). Mass transfer of all toxins was greatest under high convection with a 400,kDa membrane. Transfer of IgG and IgM was insignificant under all conditions. Bilirubin transfer was increased under high albumin conditions (P,=,0.055). Fouling of membranes ranged from 7% (400,kDa), 24% (150,kDa) to 62% (70,kDa) during a 2-h test interval. In conclusion, optimal toxin removal was achieved under high convection with a 400-kDa membrane, a condition which should provide adequate immunoprotection of hepatocytes in the BAL. Biotechnol. Bioeng. 2009; 104: 995,1003. © 2009 Wiley Periodicals, Inc. [source] Prediction of uptake dynamics of persistent organic pollutants by bacteria and phytoplanktonENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 10 2002Sabino Del Vento Abstract Phytoplankton and bacteria play an important role on the biogeochemical cycles of persistent organic pollutants (POPs). However, experimental data and quantitative knowledge of the kinetics of uptake and depuration of most POPs by bacteria and phytoplankton are scarce. In the present paper, a procedure to predict the sorption kinetics to bacteria and phytoplankton is developed. The prediction method is the combination of a mechanistic model for sorption and quantitative structure,activity relationships relating bioconcentration factors and membrane permeability to the chemical physical-chemical properties. The model consists of two compartments where the first compartment is the cellular surface and the second compartment is the cell biomass or matrix. Equations for estimating uptake and depuration rate constants into the matrix and adsorption and desorption rate constants onto the surface are obtained. These expressions depend on the physical-chemical properties of the chemical, the environmental temperature, the microorganism size, and species-specific quality of organic matter. While microorganism shape has a secondary influence on uptake dynamics, microorganism size and chemical hydrophobicity arise as the key factors controlling the kinetics of POP incorporation into bacteria and plankton. Uptake, depuration, adsorption, and desorption rate constants are reported for POPs such as polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated dioxins and furans (PCDD/Fs), and POPs of emerging concern, such as polybrominated diphenyl ethers (PBDEs). Finally, implications of uptake and depuration dynamics on the biogeochemical cycling of POPs are discussed. [source] In vitro effects of lidocaine on the contractility of equine jejunal smooth muscle challenged by ischaemia-reperfusion injuryEQUINE VETERINARY JOURNAL, Issue 1 2010M. GUSCHLBAUER Summary Reasons for performing study: Post operative ileus (POI) in horses is a severe complication after colic surgery. A commonly used prokinetic drug is lidocaine, which has been shown to have stimulatory effects on intestinal motility. The cellular mechanisms through which lidocaine affects smooth muscle activity are not yet known. Objectives: To examine the effects of lidocaine on smooth muscle in vitro and identify mechanisms by which it may affect the contractility of intestinal smooth muscle. Hypothesis: Ischaemia and reperfusion associated with intestinal strangulation can cause smooth muscle injury. Consequently, muscle cell functionality and contractile performance is decreased. Lidocaine can improve basic cell functions and thereby muscle cell contractility especially in ischaemia-reperfusion-challenged smooth muscle. Methods: To examine the effects of lidocaine on smooth muscle function directly, isometric force performance was measured in vitro in noninjured and in vivo ischaemia-reperfusion injured smooth muscle tissues. Dose-dependent response of lidocaine was measured in both samples. To assess membrane permeability as a marker of basic cell function, release of creatine kinase (CK) was measured by in vitro incubations. Results: Lidocaine-stimulated contractility of ischaemia-reperfusion injured smooth muscle was more pronounced than that of noninjured smooth muscle. A 3-phasic dose-dependency was observed with an initial recovery of contractility especially in ischaemia-reperfusion injured smooth muscle followed by a plateau phase where contractility was maintained over a broad concentration range. CK release was decreased by lidocaine. Conclusion: Lidocaine may improve smooth muscle contractility and basic cell function by cellular repair mechanisms which are still unknown. Improving contractility of smooth muscle after ischaemia-reperfusion injury is essential in recovery of propulsive intestinal motility. Potential relevance: Characterisation of the cellular mechanisms of effects of lidocaine, especially on ischaemia-reperfusion injured smooth muscle, may lead to improved treatment strategies for horses with POI. [source] Resveratrol modulates apoptosis and oxidation in human blood mononuclear cellsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 9 2003G. A. Losa Abstract Background, We examined the effect of resveratrol (RS), a nonflavonoid polyphenolic phytoalexin found in grapes and red wine, and RS coincubated with the oxidant 2-deoxy-D-ribose (dR), on apoptosis and on the oxidative metabolic status of normal human peripheral blood mononuclear cells (PBMNCs) isolated ex vivo from healthy donors. Material and methods, Apoptosis was measured by changes of membrane permeability to propidium iodide (PI), plasma membrane exposure of phosphatidylserine (PS) and intracellular caspase activity. Oxidative status was assessed by recording the intracellular glutathione concentration (GSH), the activities of the enzymes y -glutamyltransferase (y- GT) and glutathione-S-transferase (GST), and intracellular lipid peroxidation (MDA). Results, Neither apoptotic nor oxidative parameters were affected by culturing PBMNCs in medium containing RS up to 20 µM for 5 days, while the frequency of cells with intermediate permeability to PI (17% ± 5) increased at 50 µM of RS. Thus resveratrol was slightly toxic, but there was little apoptosis in these cells. Peripheral blood mononuclear cells were also grown first in medium plus RS for 24 h and then for 96 h in medium containing RS plus 10 mM of dR, an oxidant sugar that is apoptogenic for human lymphocytes. The apoptotic changes triggered by dR were counteracted by the phytoalexin in a dose-dependent manner, but RS activity was absent at the lowest concentration (5 µM) and significantly reduced at the highest concentration used (50 µM). In PBMNCs coincubated with 20 µM of RS and 10 mM of dR the antioxidant effect of RS manifested with a significant reduction of caspases-3, -8, y- GT, GST activities and MDA content. Conclusions, Peripheral blood mononuclear cells acquire antioxidant capacity when treated with RS. Grape resveratrol may make a useful dietary supplement for minimizing oxidative injury in immune-perturbed states and human chronic degenerative diseases. [source] Drug metabolism and disposition in childrenFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 3 2003M. Strolin Benedetti Abstract Key factors undergoing maturational changes accounting for differences in drug metabolism and disposition in the pediatric population compared with adults are reviewed. Gastric and duodenal pH, gastric emptying time, intestinal transit time, bacterial colonization and probably P-glycoprotein are important factors for drug absorption, whereas key factors explaining differences in drug distribution between the pediatric population and adults are membrane permeability, plasma protein concentration and plasma protein characteristics, endogenous substances in plasma, total body and extracellular water, fat content, regional blood flow and probably P-glycoprotein, mainly that present in the gut, liver and brain. As far as drug metabolism is concerned, important differences have been found in the pediatric population compared with adults both for phase I enzymes [oxidative (e.g. cytochrome CYP3A7 vs. CYP3A4 and CYP1A2), reductive and hydrolytic enzymes] and phase II enzymes (e.g. N -methyltransferases and glucuronosyltransferases). Finally, key factors undergoing maturational changes accounting for differences in renal excretion in the pediatric population compared with adults are glomerular filtration and tubular secretion. It would be important to generate information on the developmental aspects of renal P-glycoprotein and of other renal transporters as done and still being done with the different isozymes involved in drug metabolism. [source] Identification and Characterization of an Organic Solvent Tolerance Gene in Helicobacter pyloriHELICOBACTER, Issue 1 2007Hung-Chuan Chiu Abstract Background:, Pre-cleaning and soaking in glutaraldehyde is the necessary procedure to disinfect endoscopes. However, some chemical-solvent-tolerant bacteria may survive after incomplete endoscopic disinfection. The goal of this study was to identify glutaraldehyde resistance-related genes in Helicobacter pylori. Materials and Methods:, ,-Zap phagemid expression library of H. pylori strain NTUH-C1 was selected with 0.1% glutaraldehyde. The minimal inhibitory concentration (MIC) of glutaraldehyde-resistant DNA fragments of H. pylori NTUH-C1 strain were determined. Imp/OstA recombinant protein was expressed, purified, and used to generate anti-Imp/OstA polyclonal antibody. Imp/ostA knockout, deletion, and complementation strains were constructed. The function of Imp/OstA was monitored by organic solvent tolerance assay, antibiotics susceptibility test, and n -phenylnapthylamine assay. Results:, Using Imp/ostA polyclonal antibody against cell lysate of wild-type and imp/ostA mutant showed that it is not essential in H. pylori. Organic solvent tolerance assay demonstrated the role of Imp/ostA in n-hexane tolerance. MIC test showed that the mutation of imp/ostA was susceptible to hydrophobic and ,-lactam antibiotics. NPN assay demonstrated that the level of outer membrane permeability was increased by 50% in mutant strain comparing to wild-type strain (p < .001). Conclusions:, We have identified an Imp/OstA protein that was associated with glutaraldehyde resistance in our clinical strain H. pylori NTUH-C1 by screening of ,-Zap expression library. Disruption of this protein results in altering membrane permeability, sensitivity to organic solvent, and susceptibility to antibiotics. [source] DROUGHT STRESS: Comparative Time Course Action of the Foliar Applied Glycinebetaine, Salicylic Acid, Nitrous Oxide, Brassinosteroids and Spermine in Improving Drought Resistance of RiceJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 5 2010M. Farooq Abstract Worldwide rice productivity is being threatened by increased endeavours of drought stress. Among the visible symptoms of drought stress, hampered water relations and disrupted cellular membrane functions are the most important. Exogenous use of polyamines (PAs), salicylic acid (SA), brassinosteroids (BRs), glycinebetaine (GB) and nitrous oxide (NO) can induce abiotic stresses tolerance in many crops. In this time course study, we appraised the comparative role of all these substances to improve the drought tolerance in rice (Oryza sativa L.) cultivar Super-Basmati. Plants were subjected to drought stress at four leaf stage (4 weeks after emergence) by maintaining soil moisture at 50 % of field capacity. Pre-optimized concentrations of GB (150 mg l,1), SA (100 mg l,1), NO (100 ,mol l,1 sodium nitroprusside as NO donor), BR (0.01 ,m 24-epibrassinolide) and spermine (Spm; 10 ,m) were foliar sprayed at five-leaf stage (5 weeks after emergence). There were two controls both receiving no foliar spray, viz. well watered (CK1) and drought stressed (CK2). There was substantial reduction in allometric response of rice, gas exchange and water relation attributes by drought stress. While drought stress enhanced the H2O2, malondialdehyde (MDA) and relative membrane permeability, foliar spray of all the chemicals improved growth possibly because of the improved carbon assimilation, enhanced synthesis of metabolites and maintenance of tissue water status. Simultaneous reduction in H2O2 and MDA production was also noted in the plants treated with these substances. Drought tolerance was sturdily associated with the greater tissue water potential, increased synthesis of metabolites and enhanced capacity of antioxidant system. Of all the chemicals, foliar spray with Spm was the most effective followed by BR. [source] Wheat Cellular Membrane Thermotolerance Under Heat StressJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 2 2010A. S. Dias Abstract Four genotypes of Triticum aestivum L. and Triticum turgidum subsp. durum chosen according to their genetic background diversity were subjected to heat stress after anthesis. Membrane permeability, lipid peroxidation and fatty acids (C14:0, C16:0, C16:1c, C16:1t, C18:0, C18:1, C18:2 and C18:3) were quantified. The estimation of the quantum yield of non-cyclic photosynthetic electron transport was used as well as a test system to further evaluate the implications on thylakoid functioning. It was found differences within bread and durum wheat species concerning the capability to cope with high temperatures at the stage of grain filling. The genotype Sever showed high thermal sensitivity concerning membrane lipid peroxidation and membrane permeability, as evaluated by the increased production of ethylene and MDA, as well as by the impact on TFA (at the middle term of grain filling). In the durum wheat genotypes, differences were also found, with TE 9306 displaying high membrane stability, with no increases on membrane permeability, MDA and ethylene content. In this way, the observed changes on TFA in this genotype might have constituted a mechanism to allow qualitative lipid changes, reflected in lower unsaturation level of membrane FAs which is a positive trait under high temperatures. [source] Enhancement of the efficacy of erythromycin in multiple antibiotic-resistant gram-negative bacterial pathogensJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2008S. Saha Abstract Aims:, To improve the efficacy of erythromycin, a hydrophobic antibiotic, against multiple antibiotic-resistant gram-negative bacterial pathogens by enhancing their outer membrane permeability. Methods and Results:, Fifty-one nonrepeat gram-negative bacterial pathogens of various genera, resistant to multiple antibiotics, including erythromycin, were selected by disc agar diffusion tests. The amphiphilic cationic steroid antibiotic, Ceragenin CSA-13, a potent permeabilizer of bacterial outer membranes, reduced the minimum inhibitory concentration of erythromycin in 92% of the bacterial pathogens selected for the test, when supplemented with erythromycin. A synergistic effect of Ceragenin CSA-13 and erythromycin in combination was also observed. Spectrofluorimetric study confirmed that Ceragenin CSA-13 acts by depolarizing the bacterial outer membrane. The toxicity of Ceragenin CSA-13 was evaluated to be insignificant by measuring ,median lethal dose' (LD50) on mouse model. Conclusions:, Ceragenin CSA-13 may be useful as an agent to make erythromycin effective against infections caused by multiple antibiotic resistant gram-negative bacteria. Significance and Impact of the Study:, The outcome of the study suggests erythromycin,Ceragenin combination as a new approach to overcome the problem associated with the rapid emergence of multi-drug-resistant pathogens. The insignificant toxicity of Ceragenin CSA-13, as found, supports the possibility of the application of this compound for human therapeutics. [source] Investigation into the effect of detergents on disinfectant susceptibility of attached Escherichia coli and Listeria monocytogenesJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2008J.T. Walton Abstract Aims:, Investigate the effect of detergent treatment on susceptibility of attached Escherichia coli and Listeria monocytogenes to subsequent disinfectant treatment. Methods and Results:, Plate counts show that E. coli attached to stainless steel surfaces became significantly more susceptible to benzalkonium chloride (BAC) after treatment with sodium alkyl sulfate (SAS) and fatty alcohol ethoxylate (FAE). No change in susceptibility was observed with Sodium dodecyl sulfate (SDS). L. monocytogenes became significantly less susceptible to BAC after treatment with SAS and SDS yet no change in susceptibility was observed with FAE. Flow cytometry using the fluoresceine propidium iodide revealed significant increases in cell membrane permeability of both organisms by SAS and FAE, although the effect was much greater in E. coli. No change was observed with SDS. Hydrophobic interaction chromatography showed that both organisms became less hydrophobic following treatment with SAS and SDS but FAE had no effect. Conclusions:, In E. coli, detergents that increase susceptibility to BAC increase membrane permeability. In L. monocytogenes, detergents that reduce susceptibility to BAC lower cell surface hydrophobicity. Significance and Impact of the Study:, Detergents can influence the sensitivity of pathogenic food borne micro-organisms to BAC. [source] Discovery, regulation, and action of the major apoptotic nucleases DFF40/CAD and endonuclease GJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 6 2005Piotr Widlak Abstract Toward the end of the 20th and beginning of the 21st centuries, clever in vitro biochemical complementation experiments and genetic screens from the laboratories of Xiaodong Wang, Shigekazu Nagata, and Ding Xue led to the discovery of two major apoptotic nucleases, termed DNA fragmentation factor (DFF) or caspase-activated DNase (CAD) and endonuclease G (Endo G). Both endonucleases attack chromatin to yield 3,-hydroxyl groups and 5,-phosphate residues, first at the level of 50,300 kb cleavage products and next at the level of internucleosomal DNA fragmentation, but these nucleases possess completely different cellular locations in normal cells and are regulated in vastly different ways. In non-apoptotic cells, DFF exists in the nucleus as a heterodimer, composed of a 45 kD chaperone and inhibitor subunit (DFF45) [also called inhibitor of CAD (ICAD-L)] and a 40 kD latent nuclease subunit (DFF40/CAD). Apoptotic activation of caspase-3 or -7 results in the cleavage of DFF45/ICAD and release of active DFF40/CAD nuclease. DFF40's nuclease activity is further activated by specific chromosomal proteins, such as histone H1, HMGB1/2, and topoisomerase II. DFF is regulated by multiple pre- and post-activation fail-safe steps, which include the requirements for DFF45/ICAD, Hsp70, and Hsp40 proteins to mediate appropriate folding during translation to generate a potentially activatable nuclease, and the synthesis in stoichiometric excess of the inhibitors (DFF45/35; ICAD-S/L). By contrast, Endo G resides in the mitochondrial intermembrane space in normal cells, and is released into the nucleus upon apoptotic disruption of mitochondrial membrane permeability in association with co-activators such as apoptosis-inducing factor (AIF). Understanding further regulatory check-points involved in safeguarding non-apoptotic cells against accidental activation of these nucleases remain as future challenges, as well as designing ways to selectively activate these nucleases in tumor cells. © 2005 Wiley-Liss, Inc. [source] Lipophilic cationic drugs increase the permeability of lysosomal membranes in a cell culture system,JOURNAL OF CELLULAR PHYSIOLOGY, Issue 1 2010Johannes Kornhuber Lysosomes accumulate many drugs several fold higher compared to their extracellular concentration. This mechanism is believed to be responsible for many pharmacological effects. So far, uptake and release kinetics are largely unknown and interactions between concomitantly administered drugs often provoke mutual interference. In this study, we addressed these questions in a cell culture model. The molecular mechanism for lysosomal uptake kinetics was analyzed by live cell fluorescence microscopy in SY5Y cells using four drugs (amantadine, amitriptyline, cinnarizine, flavoxate) with different physicochemical properties. Drugs with higher lipophilicity accumulated more extensively within lysosomes, whereas a higher pKa value was associated with a more rapid uptake. The drug-induced displacement of LysoTracker was neither caused by elevation of intra-lysosomal pH, nor by increased lysosomal volume. We extended our previously developed numerical single cell model by introducing a dynamic feedback mechanism. The empirical data were in good agreement with the results obtained from the numerical model. The experimental data and results from the numerical model lead to the conclusion that intra-lysosomal accumulation of lipophilic xenobiotics enhances lysosomal membrane permeability. Manipulation of lysosomal membrane permeability might be useful to overcome, for example, multi-drug resistance by altering subcellular drug distribution. J. Cell. Physiol. 224:152,164, 2010 © 2010 Wiley-Liss, Inc. [source] Onion Cells After High Pressure and Thermal Processing: Comparison of Membrane Integrity Changes Using Different Analytical Methods and Impact on Tissue TextureJOURNAL OF FOOD SCIENCE, Issue 7 2010Maria E. Gonzalez Abstract:, Two different analytical methods were evaluated for their capacity to provide quantitative information on onion cell membrane permeability and integrity after high pressure and thermal processing and to study the impact of these processing treatments on cell compartmentalization and texture quality. To determine changes in cell membrane permeability and/or integrity the methodologies utilized were: (1) measurement of a biochemical product, pyruvate, formed as a result of membrane permeabilization followed by enzymatic activity and (2) leakage of electrolytes into solution. These results were compared to previously determined methods that quantified cell viability and 1H-NMR T2 of onions. These methods allowed for the monitoring of changes in the plasma and tonoplast membranes after high pressure or thermal processing. High pressure treatments consisted of 5 min holding times at 50, 100, 200, 300, or 600 MPa. Thermal treatments consisted of 30 min water bath exposure to 40, 50, 60, 70, or 90 °C. There was strong agreement between the methods in the determination of the ranges of high pressure and temperature that induce changes in the integrity of the plasma and tonoplast membranes. Membrane rupture could clearly be identified at 300 MPa and above in high pressure treatments and at 60 °C and above in the thermal treatments. Membrane destabilization effects could already be visualized following the 200 MPa and 50 °C treatments. The texture of onions was influenced by the state of the membranes and was abruptly modified once membrane integrity was lost. Practical Application:, In this study, we used chemical, biochemical, and histological techniques to obtain information on cell membrane permeability and onion tissue integrity after high pressure and thermal processing. Because there was strong agreement between the various methods used, it is possible to implement something relatively simple, such as ion leakage, into routine quality assurance measurements to determine the severity of preservation methods and the shelf life of processed vegetables. [source] The role of ultrasound and magnetic resonance in local drug deliveryJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 2 2008Roel Deckers MS Abstract Local drug delivery has recently attracted much attention since it represents a strategy to increase the drug concentration at the target location and decrease systemic toxicity effects. Ultrasound can be used in different ways to trigger regional drug delivery. It can cause the local drug release from a carrier vehicle and the local increase of cell membrane permeability either by a mechanical action or by a temperature increase. Ultrasound contrast agents may enhance these effects by means of cavitation. Ultrasound can be focused deep inside the body into a small region with dimensions on the order of 1 mm. Several types of drug microcarriers have been proposed, from nano- to micrometer sized particles. The objective of real-time imaging of local drug delivery is to assure that the delivery takes place in the target region, that the drug concentration and the resulting physiological reaction are sufficient, and to intervene if necessary. Ultrasound and nuclear imaging techniques play an important role. MRI is rather insensitive but allows precise targeting of (focused) ultrasound, can provide real-time temperature maps, and gives access to a variety of imaging biomarkers that may be used to assess drug action. Examples from recent articles illustrate the potential of the principles of ultrasound-triggered local drug delivery. J. Magn. Reson. Imaging 2008;27:400,409. © 2008 Wiley-Liss, Inc. [source] Cellular uptake and biological activity of peptide nucleic acids conjugated with peptides with and without cell-penetrating abilityJOURNAL OF PEPTIDE SCIENCE, Issue 1 2010Yvonne Turner Abstract A 12-mer peptide nucleic acid (PNA) directed against the nociceptin/orphanin FQ receptor mRNA was disulfide bridged with various peptides without and with cell-penetrating features. The cellular uptake and the antisense activity of these conjugates were assessed in parallel. Quantitation of the internalized PNA was performed by using an approach based on capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). This approach enabled a selective assessment of the PNA moiety liberated from the conjugate in the reducing intracellular environment, thus avoiding bias of the results by surface adsorption. The biological activity of the conjugates was studied by an assay based on the downregulation of the nociceptin/orphanin FQ receptor in neonatal rat cardiomyocytes (CM). Comparable cellular uptake was found for all conjugates and for the naked PNA, irrespective of the cell-penetrating properties of the peptide components. All conjugates exhibited a comparable biological activity in the 100 nM range. The naked PNA also exhibited extensive antisense activity, which, however, proved about five times lower than that of the conjugates. The found results suggest cellular uptake and the bioactivity of PNA-peptide conjugates to be not primarily related to the cell-penetrating ability of their peptide components. Likewise from these results it can be inferred that the superior bioactivity of the PNA-peptide conjugates in comparison with that of naked PNA rely on as yet unknown factors rather than on higher membrane permeability. Several hints point to the resistance against cellular export and the aggregation propensity combined with the endocytosis rate to be candidates for such factors. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd. [source] Different mechanisms of action of antimicrobial peptides: insights from fluorescence spectroscopy experiments and molecular dynamics simulations,JOURNAL OF PEPTIDE SCIENCE, Issue 9 2009Gianfranco Bocchinfuso Abstract Most antimicrobial peptides exert their activity by interacting with bacterial membranes, thus perturbing their permeability. They are investigated as a possible solution to the insurgence of bacteria resistant to the presently available antibiotic drugs. However, several different models have been proposed for their mechanism of membrane perturbation, and the molecular details of this process are still debated. Here, we compare fluorescence spectroscopy experiments and molecular dynamics (MD) simulations regarding the association with lipid bilayers and lipid perturbation for two different amphiphilic helical antimicrobial peptides, PMAP-23 and trichogin GA IV. PMAP-23, a cationic peptide member of the cathelicidin family, is considered to induce membrane permeability according to the Shai-Matsuzaki-Huang "carpet" model, while trichogin GA IV is a neutral peptide, member of the peptaibol family. Although several lines of evidence suggest a "barrel-stave" mechanism of pore formation for the latter peptide, its length is only half the normal thickness of a lipid bilayer. Both fluorescence spectroscopy experiments and MD simulations indicated that PMAP-23 associates with membranes close to their surface and parallel to it, and in this arrangement it causes a severe perturbation to the bilayer, both regarding its surface tension and lipid order. By contrast, trichogin GA IV can undergo a transition from a surface-bound state to a transmembrane orientation. In the first arrangement, it does not cause any strong membrane perturbation, while in the second orientation it might be able to span the bilayer from one side to the other, despite its relatively short length, by causing a significant thinning of the membrane. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd. [source] Membrane permeability and antimicrobial kinetics of cecropin P1 against Escherichia coli,JOURNAL OF PEPTIDE SCIENCE, Issue 6 2009Steven Arcidiacono Abstract The interaction of cecropin P1 (CP1) with Escherichiacoli was investigated to gain insight into the time-dependent antimicrobial action. Biophysical characterizations of CP1 with whole bacterial cells were performed using both fluorescent and colorimetric assays to investigate the role of membrane permeability and lipopolysaccharide (LPS) binding in lytic behavior. The kinetics of CP1 growth inhibition assays indicated a minimal inhibitory concentration (MIC) of 3 µM. Bactericidal kinetics at the MIC indicated rapid killing of E.coli (<30 min). Membrane permeability studies illustrated permeation as a time-dependent event. Maximum permeability at the MIC occurred within 30 min, which correlates to the bactericidal action. Further investigation showed that the immediate permeabilizing action of CP1 is concentration-dependent, which correlates to the concentration-dependent nature of the inhibition assays. At the MIC and above, the immediate permeability was significant enough that the cells could not recover and exhibit growth. Below the MIC, immediate permeability was evident, but the level was insufficient to inhibit growth. Dansyl polymyxin B displacement studies showed LPS binding is essentially the same at all concentrations investigated. However, it does appear that only the immediate interaction is important, because binding continued to increase over time beyond cell viability. Our studies correlated CP1 bactericidal kinetics to membrane permeability suggesting CP1 concentration-dependent killing is driven by the extent of the immediate permeabilizing action of the peptide. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd. [source] Liposome transport of hydrophobic drugs: Gel phase lipid bilayer permeability and partitioning of the lactone form of a hydrophobic camptothecin, DB-67JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 1 2008Vijay Joguparthi Abstract The design of liposomal delivery systems for hydrophobic drug molecules having improved encapsulation efficiency and enhanced drug retention would be highly desirable. Unfortunately, the poor aqueous solubility and high membrane binding affinity of hydrophobic drugs necessitates extensive validation of experimental methods to determine both liposome loading and permeability and thus the development of a quantitative understanding of the factors governing the encapsulation and retention/release of such compounds has been slow. This report describes an efflux transport method using dynamic dialysis to study the liposomal membrane permeability of hydrophobic compounds. A mathematical model has been developed to calculate liposomal membrane permeability coefficients of hydrophobic compounds from dynamic dialysis experiments and partitioning experiments using equilibrium dialysis. Also reported is a simple method to study the release kinetics of liposome encapsulated camptothecin lactone in plasma by comparing the hydrolysis kinetics of liposome entrapped versus free drug. DB-67, a novel hydrophobic camptothecin analogue has been used as a model permeant to validate these methods. Theoretical estimates of DB-67 permeability obtained from the bulk solubility diffusion model and the "barrier-domain" solubility diffusion model are compared to the experimentally observed value. The use of dynamic dialysis in drug release studies of liposome and other nanoparticle formulations is further discussed and experimental artifacts that can arise without adequate validation are illustrated through simulations. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:400,420, 2008 [source] Cerebral kinetics of oxycodone in conscious sheepJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 8 2006Hanne H. Villesen Abstract Oxycodone is an opioid analgesic that is administered orally or parenterally. The time-course of opioid action is a function of the systemic kinetics of the opioid, and the rate and extent of its entry into the brain and central nervous system. The latter is incompletely understood for oxycodone. Therefore, the cerebral kinetics of oxycodone was quantified using a conscious chronically instrumented sheep preparation. Five sheep were administered oxycodone as intravenous infusions (30 mg over 4 min). Using hybrid physiologically based kinetic models, cerebral kinetics was estimated from arterio-sagittal sinus concentration gradients and cerebral blood flow (CBF). A two-compartment membrane-limited model best described the data. The volume of the first brain compartment was 35.4 mL with a half-life of equilibrium of 0.6 min. The brain:blood equilibration of oxycodone was relatively slow (half-life of 7.2 min), with a large deep cerebral distribution volume (222.8 mL) for the second compartment and a moderate membrane permeability of 54.8 mL/min, which exceeded the nominal CBF (40 mL/min). Drug retention in the brain was 1.3% after 45 min. In conclusion, pharmacokinetic modelling of oxycodone showed a delayed equilibration between brain and blood of a nature that would be affected by changes in both CBF and blood brain barrier permeability. © 2006 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 95: 1666,1676, 2006 [source] QSAR analysis of interstudy variable skin permeability based on the "latent membrane permeability" conceptJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 10 2003Shin-Ichi Fujiwara Abstract A number of QSAR models for skin permeability have been proposed, but these models lack consistency due to interspecies and interlaboratory differences. This study was initiated to extract an essential QSAR from the multiplicity of data sets of skin permeability by using a novel statistical approach. Ten data sets were collected from the literature, which include a total of 111 permeability coefficients in human, hairless mouse, or hairless rat skin for 94 structurally diverse compounds. Following a Potts and Guy's approach, the octanol/water partition coefficient and molecular weight were chosen as molecular descriptors. All of the data sets were analyzed simultaneously, assuming that all of the sets share a latent, common factor as far as the structure/permeability relationship is concerned. Despite the fact that the degree-of-freedom for the present analysis was limited compared with that for individual regression analyses, the determination coefficients (R2) were high enough for all the 10 data sets, with an average R2 of 0.815 (average R2,=,0.825 for individual analyses). Thus, skin permeability of compounds can be well explained from the log P and M.W., where the ratio of the contribution to skin permeability was approximately 1:1. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 92:1939,1946, 2003 [source] Permeability of a novel ,-lactamase inhibitor LK-157 and its ester prodrugs across rat jejunum in vitroJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 9 2009Petra Igli Abstract Objectives LK-157 is a novel 10-ethylidene tricyclic carbapenem that resembles the structure of the broad-spectrum antibiotic sanfetrinem and acts as a potent inactivator of ,-lactamases of classes A, C and D. LK-157 is a highly soluble but poorly permeable drug. Since most of the ,-lactams are poorly absorbed, ester prodrugs LK-159, LK-157E1 and LK-157E2 were designed to enhance membrane permeability. This study investigated the permeability of LK-157 and the three ester prodrugs across rat intestine in vitro. The morpholinoethyl ester of sanfetrinem was also investigated. Method Permeability across rat jejunum was determined using EasyMount side-by-side diffusion chambers. Key findings The solubility and permeability of morpholinoethyl ester LK-157E2 were superior to those of LK-159 and LK-157E1. The morpholinoethyl ester of sanfetrinem LK-176E1 had the highest observed permeability coefficient and consequently the highest predicted absorption in humans. Conclusions These results suggest that the morpholinoethyl esters of LK-157 and sanfetrinem could be further investigated to assess bioavailability in vivo. [source] Impact of hot water treatment on sprouting, membrane permeability, sugar content and chip colour of reconditioned potato tubers following long-term cold storageJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 15 2008Marios C Kyriacou Abstract BACKGROUND: The efficacy of hot water treatment in facilitating successful reconditioning of processing potato (Solanum tuberosum L.) cultivar Hermes following 6 months cold storage at 4.5 °C was examined. Tubers were subjected to hot water treatments (HWTs) at 52.5, 55.0, 57.5 and 60.0 °C for 0,60, 0,50, 0,40 and 0,20 min, respectively, and then reconditioned for 20 days at 16 °C before evaluated for sprouting, fresh weight loss, membrane permeability, sugar content and processing quality. RESULTS: The study demonstrates that in order to achieve complete inhibition of sprouting during potato reconditioning HWTs must exceed the thermal tolerance threshold of the tubers. Short-duration HWT was effective in retarding sprout growth and tuber dehydration without significantly affecting storage parenchyma membrane permeability, tuber sugar content or processing quality. On the contrary, prolonged HWT caused extensive heat damage, loss of membrane integrity and induced an increase in tuber sucrose and reducing sugar content resulting in deterioration of chip colour in proportion to treatment duration. CONCLUSION: Although HWT at 52.5,60 °C following long-term cold storage did not improve the processing quality of potato tubers after 20 days of reconditioning, future work is needed to evaluate the effect of short-duration HWT on the permissible extent of reconditioning and subsequent processing quality. Copyright © 2008 Society of Chemical Industry [source] Morphological changes in mouse embryos cryopreserved by different techniquesMICROSCOPY RESEARCH AND TECHNIQUE, Issue 4 2007A.R.S. Coutinho Abstract Cryopreservation of mammalian embryos is an important tool for the application of reproductive biotechnologies. Subjective evaluation to determine embryo viability is often used. The determination of the best cryopreservation protocol depends on morphological and molecular analysis of cellular injuries. The main objective of this study was to compare two methods of cryopreservation by assessing morphological alterations of frozen embryos using light, fluorescence, and transmission electron microscope. Fresh (control), slow frozen, and vitrified mouse embryos were composed. To evaluate the viability of the embryos, the cell membrane integrity was assessed using Hoechst33342 and propidium iodide (H/PI) staining. Morphological analyses using hematoxylin and eosin (HE) staining were performed to test different techniques (in situ, paraffin, and historesin) by both light and fluorescence microscopy. Transmission electron microscope was used to detect ultrastructural alterations in Spurr- and Araldite-embedded samples. H/PI staining detected more membrane permeability in the vitrification (69.8%) than in the slow freezing (48.4%) or control (13.8%) groups (P < 0.001). Historesin-embedded samples showed to be more suitable for morphological analyses because cellular structures were better identified. Nuclear evaluation in historesin sections showed the induction of pycnosis in slow freezing and vitrification groups. Cytoplasm evaluation revealed a condensation and an increase in eosinophilic intensity (indicating apoptosis) in the slow freezing group, and weakly eosinophilic structures and degenerated cells (indicating oncosis) in the vitrification group (P < 0.05). Ultrastructural analyses confirmed HE morphological findings. It was concluded that both cryopreservation techniques resulted in oncosis and apoptosis injuries. However, vitrification caused more severe cellular alterations and reduced embryonic viability compared to slow freezing. Microsc. Res. Tech., 2007. © 2006 Wiley-Liss, Inc. [source] Maintaining a healthy SPANC balance through regulatory and mutational adaptationMOLECULAR MICROBIOLOGY, Issue 1 2005Thomas Ferenci Summary Stress protection is an important but costly contributor to bacterial survival. Two distinct forms of environmental protection share a common cost and a significant species-wide variability. Porin-mediated outer membrane permeability and the RpoS-controlled general stress response both involve a trade-off between self- preservation and nutritional competence, called the SPANC balance. Interestingly, different Escherichia coli strains exhibit distinct settings of the SPANC balance. It is tilted towards high stress resistance and a restricted diet in some isolates whereas others have broader nutritional capability and better nutrient affinity but lower levels of resistance. Growth- or stress-related selective pressures working in opposite directions (antagonistic pleiotropy) result in polymorphisms affecting porins and RpoS. Consequently, these important cellular components are present at distinct concentrations in different isolates. A generalized hypothesis to explain bacterial adaptation, based on the SPANC investigations, is offered. A holistic approach to bacterial adaptation, involving a gamut of regulation and mutation, is likely to be the norm in broadening the capabilities of a species. Indeed, there is unlikely to be a standard regulatory setting typical for all members of a species. Gene regulation provides a limited fine control for maintaining the right level of adaptation in a particular niche but mutational changes provide the coarse control for adaptation between the species-wide environments of free-living bacteria. [source] Free Zn2+ enhances inhibitory effects of EGCG on the growth of PC-3 cellsMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 4 2008Shi-li Sun Abstract Epigallocatechin-3-gallate (EGCG), a major component of green tea, has both preventive and therapeutic beneficial actions in prostate cancer. In the present study, we compared the growth inhibitory effects and the antioxidant and ability to modify cell membrane permeation of zinc-EGCG complex and Zn2+/EGCG mixture on androgen-insensitive prostate cancer (PC-3) cells. It was noted that free Zn2+ enhanced the growth inhibitory effects of EGCG on PC-3 cells at 160 ,mol/L concentration, whereas zinc-EGCG complex was ineffective. EGCG showed potent free radical scavenging ability in the presence of Zn2+. EGCG in the presence of Zn2+ was more effective than EGCG alone in enhancing the permeability of the cell membrane, whereas zinc-EGCG complex had no effect on PC-3 cell membrane permeability. These results indicate that though Zn2+ enhanced the action of EGCG on PC-3 cells, zinc-EGCG complex is highly unlikely to be formed in the presence of Zn2+ and EGCG to explain the potentiating action of Zn2+ on the growth inhibitory property of EGCG on PC-3 cells. [source] Two daytime icodextrin exchanges decrease brain natriuretic peptide levels and improve cardiac functions in continuous ambulatory peritoneal dialysis patientsNEPHROLOGY, Issue 3 2010TANSU SAV ABSTRACT Aim: Peritoneal dialysis patients with ultrafiltration failure frequently have fluid overload. It is known that the increase in the ultrafiltration is associated with decrease in the left ventricle (LV) dysfunction. This study was designed to examine the potential effects of serum brain natriuretic peptide (BNP) on cardiac functions and to determine the relationship between BNP and cardiac parameters in continuous ambulatory peritoneal dialysis (CAPD) patients with ultrafiltration failure. Methods: Twenty-eight patients with high or high-average membrane permeability as indicated by the peritoneal equilibration test were enrolled and randomized to receive either once or twice daily icodextrin. Serum BNP levels and echocardiographic measurements were evaluated at baseline and at the end of the eighth week. The correlations between the percentage changes of parameters from baseline were also studied. Results: In both groups there was a significant decrease in serum BNP, LV mass, heart rate (HR) and cardiothoracic index (CTI) and an improvement in ejection fraction (all P < 0.05). However, the percentage of change in all these parameters was significantly better in the twice daily compared with once daily group (all P < 0.05). Furthermore, the percentage decrease in BNP was positively correlated with the percentage decrease in HR, LV mass and BP. Conclusion: Twice daily icodextrin treatment might be useful in hypervolaemic CAPD patients for the improvement of cardiac functions. BNP monitoring may be useful to follow up these patients. [source] | |||||||||||||||||||||||||||||||