The Experts below are selected from a list of 1635 Experts worldwide ranked by ideXlab platform

Eric Bakker - One of the best experts on this subject based on the ideXlab platform.

  • equipment free detection of k on microfluidic paper based analytical devices based on exhaustive replacement with ionic dye in ion selective capillary sensors
    ACS Sensors, 2019
    Co-Authors: Yoshiki Soda, Daniel Citterio, Eric Bakker
    Abstract:

    A distance-based analysis of Potassium ion (K+) is introduced that is performed on a microfluidic paper-based analytical device (μPAD) coupled to an ion-selective capillary sensor. The concept is based on two sequential steps, the selective replacement of analyte ion with an ionic dye, and the detection of this dye in a distance-based readout on paper. To achieve the first step, the capillary sensor holds a poly(vinyl chloride) (PVC) membrane film layer plasticized by dioctyl sebacate (DOS) that contains the Potassium Ionophore valinomycin, a lipophilic cation-exchanger and the ionic indicator Thioflavin T (ThT) on its inner wall. Upon introduction of the sample, K+ in the aqueous sample solution is quantitatively extracted into the film membrane and replaced with ThT. To convert the ion exchange signal into a distance-based analysis, this solution was dropped onto the inlet area of a μPAD to flow the ThT along a channel defined by wax printing, resulting in the electrostatic binding of ThT to the cellulo...

  • Equipment-Free Detection of K+ on Microfluidic Paper-Based Analytical Devices Based on Exhaustive Replacement with Ionic Dye in Ion-selective Capillary Sensors
    2019
    Co-Authors: Yoshiki Soda, Daniel Citterio, Eric Bakker
    Abstract:

    A distance-based analysis of Potassium ion (K+) is introduced that is performed on a microfluidic paper-based analytical device (μPAD) coupled to an ion-selective capillary sensor. The concept is based on two sequential steps, the selective replacement of analyte ion with an ionic dye, and the detection of this dye in a distance-based readout on paper. To achieve the first step, the capillary sensor holds a poly­(vinyl chloride) (PVC) membrane film layer plasticized by dioctyl sebacate (DOS) that contains the Potassium Ionophore valinomycin, a lipophilic cation-exchanger and the ionic indicator Thioflavin T (ThT) on its inner wall. Upon introduction of the sample, K+ in the aqueous sample solution is quantitatively extracted into the film membrane and replaced with ThT. To convert the ion exchange signal into a distance-based analysis, this solution was dropped onto the inlet area of a μPAD to flow the ThT along a channel defined by wax printing, resulting in the electrostatic binding of ThT to the cellulose carboxylic groups. The initial amount of K+ determines the amount of ThT in the aqueous solution after ion-exchange, and consequently the distance of ThT-colored area reflects the sample K+ concentration. The ion exchange reaction was operated in a so-called “exhaustive sensing mode” and gave a distinct response in a narrow range of K+ concentration (1–6 mM) that cannot be achieved by the classical optode sensing mode. The absence of hydrogen ions from the equilibrium competition of the capillary sensor contributed to a complete pH-independence, unlike conventional optodes that contain a pH sensitive indicator. A very high selectivity for K+ over Na+ and Ca2+ has been confirmed in separate solutions and mixed solutions tests. K+ measurements in pooled serum samples at concentrations between 2 and 6 mM are successfully demonstrated on a temperature controlled support

  • Selective Distance-Based K+ Quantification on Paper-Based Microfluidics
    2018
    Co-Authors: Chase T. Gerold, Eric Bakker, Charles S. Henry
    Abstract:

    In this study, paper-based microfluidic devices (μPADs) capable of K+ quantification in aqueous samples, as well as in human serum, using both colorimetric and distance-based methods are described. A lipophilic phase containing Potassium Ionophore I (valinomycin) was utilized to achieve highly selective quantification of K+ in the presence of Na+, Li+, and Mg2+ ions. Successful addition of a suspended lipophilic phase to a wax printed paper-based device is described and offers a solution to current approaches that rely on organic solvents, which damage wax barriers. The approach provides an avenue for future alkali/alkaline quantification utilizing μPADs. Colorimetric spot tests allowed for K+ quantification from 0.1–5.0 mM using only 3.00 μL of sample solution. Selective distance-based quantification required small sample volumes (6.00 μL) and gave responses sensitive enough to distinguish between 1.0 and 2.5 mM of sample K+. μPADs using distance-based methods were also capable of differentiating between 4.3 and 6.9 mM K+ in human serum samples. Distance-based methods required no digital analysis, electronic hardware, or pumps; any steps required for quantification could be carried out using the naked eye

  • multiplexed flow cytometric sensing of blood electrolytes in physiological samples using fluorescent bulk optode microspheres
    Analytical Chemistry, 2007
    Co-Authors: Katarzyna Wygladacz, Robert Retter, Michael Bell, Eric Bakker
    Abstract:

    Polymeric bulk optode microsphere ion sensors in combination with suspension array technologies such as analytical flow cytometry may become a power tool for measuring electrolytes in physiological samples. In this work, the methodology for the direct measurement of common blood electrolytes in physiological samples using bulk optode microsphere sensors was explored. The simultaneous determination of Na+, K+, and Ca2+ in diluted sheep blood plasma was demonstrated for the first time, using a random suspension array containing three types of mixed microsphere bulk optodes of similar size, fabricated from the same chromoIonophore without additional labeling. Sodium Ionophore X, Potassium Ionophore III, and grafted AU-1 in poly(butyl acrylate) were the Ionophores used in the bulk optode microsphere ion sensors for Na+, K+, and Ca2+, respectively, in combination with the cation-exchanger NaTFPB (sodium tetrakis-[3,5-bis(trifluoromethyl)phenyl]borate) and the same concentration of the chromoIonophore ETH 5294 ...

Hansdietmar Beer - One of the best experts on this subject based on the ideXlab platform.

  • genome editing of human primary keratinocytes by crispr cas9 reveals an essential role of the nlrp1 inflammasome in uvb sensing
    Journal of Investigative Dermatology, 2018
    Co-Authors: Gabriele Fenini, Serena Grossi, Thomas Biedermann, Ernst Reichmann, Emmanuel Contassot, Lars E French, Hansdietmar Beer
    Abstract:

    By forming a protective barrier, epidermal keratinocytes represent the first line of defense against environmental insults. UVB radiation of the sun is a major challenge for the skin and can induce inflammation, aging, and eventually skin cancer. UVB induces an immune response in human keratinocytes resulting in activation and secretion of the proinflammatory cytokines proIL-1β and -18. This is mediated by an assembly of protein complexes, termed inflammasomes. However, the mechanisms underlying sensing of UVB by keratinocytes, and particularly the types of inflammasomes required for cytokine secretion, are a matter of debate. To address these questions, we established a protocol that allows the generation of CRISPR/Cas9-targeted human primary keratinocytes. Our experiments showed an essential role of the NLRP1 rather than the NLRP3 inflammasome in UVB sensing and subsequent IL-1β and -18 secretion by keratinocytes. Moreover, NLRP1 but not NLRP3 was required for inflammasome activation in response to nigericin, a Potassium Ionophore and well-established NLRP3 activator in immune cells. Because the CRISPR/Cas9-targeted cells retained their full differentiation capacity, genome editing of human primary keratinocytes might be useful for numerous research and medical applications.

Ikuko Ueno - One of the best experts on this subject based on the ideXlab platform.

  • induction of apoptosis by t 2 toxin and other natural toxins in hl 60 human promyelotic leukemia cells
    Natural Toxins, 1995
    Co-Authors: Yoshio Ueno, Kiyoko Umemori, Eeichi C Niimi, Seiichi Tanuma, Satoshi Nagata, Masao Sugamata, Tomomi Ihara, Masaru Sekijima, Kenichi Kawai, Ikuko Ueno
    Abstract:

    Based on the DNA fragmentation profile in gel electrophoresis and the morphological changes in electron microscopy, the induction of apoptotic nuclear changes by mycotoxins and other microbial products, in total 31 chemicals, was investigated in HL-60 human promyelotic leukemia cells, along with the cytotoxicity tests with 3-[4,5-dimethylthiazol-zyl]-2,5-diphenyltetrazolium bromide (MTT) and trypan blue exclusion. Among the chemicals tested, trichothecenes (T-2 toxin, roridin A, nivalenol, deoxynivalenol), certain anthraquinones (luteoskyrin, skyrin, 2-hydroxyemodin), diketopiperazines (emethallicin A, emestrin), isocoumarins (ochratoxin A, citrinin), lactone (penicillic acid), dihydrobisfuran (aflatoxin B), Potassium Ionophore (valinomycin), and an inhibitor of interleukin-2 synthesis (cyclosporin A) were positive for the induction of DNA fragmentation. No DNA fragmentation was observed under the present conditions with fumonisin B1, cyclic peptides (cyclochlorotine, phalloidin, microcystin-LR), certain anthraquinones (emodin, chrysophanol, rugulosin), and others (sterigmatocystin, cytochalasin A, griseofulvin, fusaric acid, kojic acid, rubratoxin B, butenolide, wortmannin, FK506, and sphingosine). The apoptotic changes in the cells exposed to T-2 toxin and luteoskyrin were confirmed by electron microscopic observation. Detailed experiments on dose and time dependencies revealed that T-2 toxin induced the apoptosis at 10 ng/ml (= 4 × 10-8 M) levels within 2-6 hr without significant cytotoxicity evaluated by the dye exclusion and MTT. © 1995 Wiley-Liss, Inc.

  • induction of apoptosis by t 2 toxin and other natural toxins in hl 60 human promyelotic leukemia cells
    Natural Toxins, 1995
    Co-Authors: Yoshio Ueno, Kiyoko Umemori, Eeichi C Niimi, Seiichi Tanuma, Satoshi Nagata, Masao Sugamata, Tomomi Ihara, Masaru Sekijima, Kenichi Kawai, Ikuko Ueno
    Abstract:

    Based on the DNA fragmentation profile in gel electrophoresis and the morphological changes in electron microscopy, the induction of apoptotic nuclear changes by mycotoxins and other microbial products, in total 31 chemicals, was investigated in HL-60 human promyelotic leukemia cells, along with the cytotoxicity tests with 3-[4,5-dimethylthiazol-zyl]-2,5-diphenyltetrazolium bromide (MTT) and trypan blue exclusion. Among the chemicals tested, trichothecenes (T-2 toxin, roridin A, nivalenol, deoxynivalenol), certain anthraquinones (luteoskyrin, skyrin, 2-hydroxyemodin), diketopiperazines (emethallicin A, emestrin), isocoumarins (ochratoxin A, citrinin), lactone (penicillic acid), dihydrobisfuran (aflatoxin B1), Potassium Ionophore (valinomycin), and an inhibitor of interleukin-2 synthesis (cyclosporin A) were positive for the induction of DNA fragmentation. No DNA fragmentation was observed under the present conditions with fumonisin B1, cyclic peptides (cyclochlorotine, phalloidin, microcystin-LR), certain anthraquinones (emodin, chrysophanol, rugulosin), and others (sterigmatocystin, cytochalasin A, griseofulvin, fusaric acid, kojic acid, rubratoxin B, butenolide, wortmannin, FK506, and sphingosine). The apoptotic changes in the cells exposed to T-2 toxin and luteoskyrin were confirmed by electron microscopic observation. Detailed experiments on dose and time dependencies revealed that T-2 toxin induced the apoptosis at 10 ng/ml (= 4 x 10(-8) M) levels within 2-6 hr without significant cytotoxicity evaluated by the dye exclusion and MTT.

Satya N Mathur - One of the best experts on this subject based on the ideXlab platform.

  • esterification of plasma membrane cholesterol and triacylglycerol rich lipoprotein secretion in caco 2 cells possible role of p glycoprotein
    Journal of Lipid Research, 1995
    Co-Authors: F J Field, E Born, Heshun Chen, Shubha Murthy, Satya N Mathur
    Abstract:

    Acylcoenzyme A:cholesterol acyltransferase (ACAT) and/or cholesteryl esters have been implicated as important factors in the normal assembly of apolipoprotein (apoB)-containing lipoproteins. The predominant substrate for ACAT is believed to originate from cholesterol contained within the plasma membrane. To investigate a possible role of intestinal plasma membrane cholesterol in triacylglycerol-rich lipoprotein synthesis and secretion, CaCo-2 cells were incubated with agents that are known to interfere with cholesterol transport from the plasma membrane to the ER. Progesterone, verapamil, and trifluoperazine significantly decreased the movement of cholesterol from plasma membrane to endoplasmic reticulum (ER) in CaCo-2 cells. Without altering the synthesis of apoB and independent of their effects on cellular cholesterol esterification, progesterone, verapamil, and trifluoperazine decreased the basolateral secretion of triacylglycerols, cholesteryl esters, and immunoreactive and newly synthesized apoB. The three agents also interfered with the esterification of cholesterol absorbed from taurocholate micelles. As progesterone, verapamil, and trifluoperazine are recognized inhibitors of p-glycoprotein, a variety of agents that have been shown to interfere with p-glycoprotein function were tested to investigate their effects on cholesterol transport and apoB secretion. All the agents significantly decreased in parallel both cholesterol transport and apoB secretion. In contrast, methotrexate, an antimetabolite that does not interact with p-glycoprotein, had no effect. Nigericin, a Potassium Ionophore, which causes alkalinization of intracellular vesicles, also caused a profound inhibition of cholesterol transport and apoB secretion. Preventing plasma membrane cholesterol from arriving at the ER, or inhibiting the esterification of plasma membrane cholesterol, does not alter apoB secretion. However, the results suggest a possible role for p-glycoprotein in normal cholesterol trafficking and triacylglycerol-rich lipoprotein secretion in CaCo-2 cells. It is postulated that p-glycoprotein might function to maintain the acidic environment of transport vesicles, and therefore, could play a role in the transport of lipids by the intestine.

Gabriele Fenini - One of the best experts on this subject based on the ideXlab platform.

  • genome editing of human primary keratinocytes by crispr cas9 reveals an essential role of the nlrp1 inflammasome in uvb sensing
    Journal of Investigative Dermatology, 2018
    Co-Authors: Gabriele Fenini, Serena Grossi, Thomas Biedermann, Ernst Reichmann, Emmanuel Contassot, Lars E French, Hansdietmar Beer
    Abstract:

    By forming a protective barrier, epidermal keratinocytes represent the first line of defense against environmental insults. UVB radiation of the sun is a major challenge for the skin and can induce inflammation, aging, and eventually skin cancer. UVB induces an immune response in human keratinocytes resulting in activation and secretion of the proinflammatory cytokines proIL-1β and -18. This is mediated by an assembly of protein complexes, termed inflammasomes. However, the mechanisms underlying sensing of UVB by keratinocytes, and particularly the types of inflammasomes required for cytokine secretion, are a matter of debate. To address these questions, we established a protocol that allows the generation of CRISPR/Cas9-targeted human primary keratinocytes. Our experiments showed an essential role of the NLRP1 rather than the NLRP3 inflammasome in UVB sensing and subsequent IL-1β and -18 secretion by keratinocytes. Moreover, NLRP1 but not NLRP3 was required for inflammasome activation in response to nigericin, a Potassium Ionophore and well-established NLRP3 activator in immune cells. Because the CRISPR/Cas9-targeted cells retained their full differentiation capacity, genome editing of human primary keratinocytes might be useful for numerous research and medical applications.