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Avi markovich affinity investments network

Gazit, A. Polymer Science , Gabashvili, Mater. Gedanken, Ultrasonic Sonochem. Rozenberg, S. Banerjee, I. Sominski and A. Pol, Vilas G. Ben-Ishay, A. Kotlyar, N. Amiryan, M. Meyer, W. Langmuir 23 , Gedanken, and Ya. Tuval and A. Pol, Aharon Gedanken, I. Felner, I. Sung, ; S. Asai, Inorg. Jung, , A.

Koltypin, Phys. Koltypin, V. Gedanken, D. Aurbach J. Irzh, N. Gedanken, Langmuir 23 , Gabashvili, Dana. B , Pol and A. Jung, Ya. Rozenberg,1, A. Auslender, G. Banerjee, D. B 76 , Solovyov, Evangelia Xenogiannopoulou,. Perkas, V. Gorodetsky, E. Gedanken and Ya. Kessler, and Aharon Gedanken, New J. Abu-Much, U. Jacob, Jimmy C. Yu, Aharon Gedanken, Chem.

Pang, G. Chem B , Jung, M. Mukovskii, Submitted to J. Augustin, C. Sanjeeviraja, V. Zhong, F. Chen, T. Ang, Y. Han, W. Lim, A. Gedanken, Inorg. B ; , Kim, D. Glass Phys. Landau, Aharon Gedanken, Langmuir 22, Pol, P. George, B. Markovsky and A. Kim, A. Tver'yanovich, D. Lee, and B.

Pol, Aharon Gedanken Carbon 44 , Turgeman and A. Gedanken, Cryst. Zhou, K. Huang, U. Frydrnan, A. Langmuir 22 , Pol, Jose M. Calderon Moreno, Aharon Gedanken, Carbon 44 , Rangari, D. Srivastava, A. Grzanka, and W. Yatsunenko, V. Osinniy and T. Story, Appl. Sivakumar, A. Bhattacharya, Y. Yeshurun, W. Zhong, Y. Du, I. Felner, J. Turgeman, O. Gershevitz, O. Palchik, M.

Deutsch, B. Ocko, A. Gedanken, C. Gofer, J. Edited by C. Hoz, L. Luong, Aharon Gedanken, J. Oliete , T. Seisenbaeva, M. Sung, S. Asai, and A. B, , Pol, Gulaim Seisenbaeva, Vadim G. Kessler, and A. Bahadur, S. Vitta, O. Prakash, Narosa Publishing House, , Pages. Nikitenko, Yu. Koltypin, I. Yeshurun, A. Shames, J. Jiang, V. Markovich, G. Gorodetsky, A. Rana, V. Pol, I.

Frydman, Adv. Gedanken, Ultrasonic Sonochemistry 11 , - , Gedanken, V. Gorodetsky, G. Pang, and A. Gedanken, B. Atkas et al. Brukental, Y. Nowik, Chem. Qiu, Vilas G. Calderon-Moreno, and Aharon Gedanken, Langmuir 20 , Kumar, J. Gnanaraj, G. Salitra, A. Abramov, A. Aurbach, J. Soupart and J. Rousche, Journal of Solid State Electrochemistry 8, - Koltypin, N.

Perkas, M. Besson, L. Vradman, M. Hershkovitz, M. Landau, Trans. Tang, W. Huang Huang, S. Liu, Y. Wang, O. Palchik, Yu. Koltypin, D. Pickup, E. Jortner 70th birthday. State Chem. Nanoparticles Research 6, Srivastava, N. Seisenbaeva, Y. Kessler and A. Van-Eck, A. Gedanken, Ultrason.

Ganesh Kumar, D. Aurbach, and A. Gedanken Ultrasonic Sonochemistry 10, 17 Landau, M. Herskowitz, V. Ezersky, M. Talianker, S. Gedanken, and J. Calderon-Moreno, Chem. Pol, M. Motiei, A. Mastai, Chem. Topaz, A. Rubowitz, E. Assia, M. Rosner, D. Meyerstein, N. Meyerstein and A. Perkas, H. Buchkremer, N. Rana, X. Xu, Y. Yeshurun and A. Palchik, A. Odani, A. Nimberger, B. Markovsky, E. Levi, V. Kumar, M. Gedanken, P. Dan, D. Aurbach, Journal of Power Sources, , Menzler, D.

Lavergnat, F. Tietz, E. Djurado, W. Fischer, G. Pang, A. Gedanken, H. New J. Gedanken, Stud. Palchik, V. Pol, E. Sominska, Yu. Perkas, O. Gedanken, Submitted to J. Gnanaraj, V. Gedanken and D. Aurbach, Electrochem. Gnanaraj, S. Ben-David, D.

Aurbach, Chem. Gabashvili, G. Seisenbaeva, V. Kessler, L. Zhang, J. Yu, A. Gedanken, Current Science, 85, No. Avivi Levi, , Y. Dror and A. Topaz, E. Assia, D. Meyerstein, A. Gedanken, Invest. Assia, A. Rubowitz, M.

Topaz, Invest. Palchik, O. Koltypin, Y. Diamant, Y. Sominska, E. Grisaru, O. Slifkin, A. Hong-liang Li, Ying-chun Zhu, O. Gedanken, I. Chem B , - Jeevanandam, Yu. B 90, Koltypin, S. Nikitenko, A. Pol, D. Srivastava, O. Gedanken, Langmuir 18, Wolfus, A.

Shaulov, Y. Yeshurun, I. Novik, Yu. Rana, A. Yeshurun, X. Xu, J. Weiss, J. Rozenberg G. Arsenov, D. Shulyatev, Ya. Mukovskii, A. Gedanken, G. Pang, J. Harpeness, O. Palchik, S. Amiel, M. Weiss, Chem. Motiei, J. Calderon-Moreno, A. Socol, O. Abramson, A. Meshorer, A. Aurbach, A. Koltypin, O. Palchik, and A. Gedanken, Biochemical Journal , Topaz, M. Meyerstein, Ultrasound in Medicine and Biology 28 , Huang, X.

Tang, I. Felner, Yu. Kumar Rana, Y. Gedanken Adv. Pol, R. Reisfeld, and A. Pol, O. Felner, Appl. Nimberger, E. Levi, E. Gedanken Chem. Stanislav Kishinevski, Sergei I. Pickup, Ernst R. Publications in Wang, S. Chen, X. Tang, O.

Zaban, Yu. Vijaya Kumar, R. Elgamiel, Y. Gedanken , Langmuir, 17 , Palchik, R. Kerner, A. Weiss, M. Slifkin, V. Palchik J. Gadanken J. Vijaya Kumar, Y. Mastai, Y. Diamant, A. Kerner, O. Pang, E. Sominska, H. Colfen, Y. Mastai, S. Avivi, Yu, Koltypin, A.

Gedanken, Nanoletters 1, Liu, W. Huang, S. Chen, S. Avivi, A. Non-Crystalline Solids , Pang, S. Chen, Y. Zhu, O. Vijayakumar, Yu. Koltypin, X. Landau, L. Vradman and M. Herskowitz, Y. Meyerstein, Ultrasound in Medicine and Biology, 27, Avivi and A. Momma, N. Shiraishi, A. Yoshizawa, T. Osaka, A. Zhu, L. Sominska, J. Power Sources , , Sp. SI Fernandez, M. Sayagues, J. Fischer, Aharon Gedanken, J.

Suwen, L. Gedanken, X. Felner, G. Irradiation, Y. Wang, L. Yin, O. Palchik, Y. Rosenfeld Hacohen, Yu. Gedanken , Langmuir 17, Rohit Kumar Rana, Yu. Avivi Levi , I. Novik and A. Jeevanandam, Y. Diamant, M. Gedanken Phys. Weiss, Y. Rozenfeld Hacohen, Inorganic Chemistry 40, Aharon Gedanken, Y.

Wang, X. Tang, N. Perkas, Yu. Koltypin, M. Felner, A. Gedanken, Angew. Chen, R. Vijaya Kumar, A. Zaban, A. Zhu, Yu. Zhu, S. Aruna, Yu. Mastai, J. Ramesh, E. Sominska, B. Cina, R. Chaim and A. Salkar, P. Jeevanandam, S. Palchik and A. Shafi, A. Prozorov, A. Revesz, J.

Mestai, R. Salkar, Y. Rojas, M. Sayagues, A. Caballero, A. Fernandez, Yu. Gedanken, L. Ponsonnet, B. Vacher, J. Martin, J. Vijaykumar, Y. Cohen, Yair Cohen, D. Palchik, J. Zhu, A. Ramesh, I. Felner, Y. Wizel, S. Margel, A. Gedanken, Polymer International. Avivi, Y. Mastai, and A. JACS , Tang, L. Yin, Yu. Vijaykumar, Yu. A , Tang, Y. Wang, Yu. Liu, O. Gedanken, Langmuir 16, Reisfeld, E. Sominski, Yu. Palchik, G. Panzer, M. Gaft, H. Suwen Liu, J. Zhu, Y. Mestai, I. Yin, W. Huang, A.

Reisfeld, L. Sominski, Z. Zhong, Yu. Koltypin, G. Panczer, M. Koltypin, and A. Zhu, Z. Lu, T. Aruna, D. Palchik, I. Kataby, T. Tang , S. Liu , Y. Wang , W. Huang Yu. Gedanken, to Chem. Gofer, Y. Kerner, Z. Zhu and A. Raz, J. Jortner and A. Alveolar fluid clearance in human lungs ex vivo. All patients were given oral and written informed consent forms. Human lung tissues were obtained during pulmonary resection surgeries with lung cancer at the First Affiliated Hospital of CMU.

There were no fibrous or emphysematous lesions as assessed by preoperative chest CT scanning. Human lung segments were prepared, and AFC measurements were done as described previously Briefly, the segmental bronchus was occluded by a Fr. Alveolar fluid was aspirated 60 minutes after instillation. In vivo alveolar fluid clearance AFC in mice. In vivo AFC rate was measured as previously described 68 - To maximize the collection of instilled BSA solution, the diaphragm was dissected.

Methodological concerns were addressed by using amiloride to inhibit water movement to confirm that measurements accurately reflected AFC. The endogenous murine albumin would lead to an overestimation of AFC rates. This potential bias was corrected by measuring murine albumin AssayPro, St. Final AFC values were corrected removing murine albumin present in the aspirate. In addition, we used an aliquot of the instillate that had been delivered into the lungs and removed within 5 min, rather than freshly prepared naive instillate per se.

Any pre-existing edema or leaking murine plasma proteins would be excluded for it dilutes both the control at time point 0 min P i and final samples P f. Mouse AT2 cell isolation and monolayer cultures. Mouse tracheal epithelial cell isolations and monolayer cultures.

Briefly, mice were anesthetized with intraperitoneal injection of ketamine HCl and xylazine. The trachea proximal to the bronchial bifurcation was isolated and removed. The resected section was immediately placed in PBS. Under a dissecting microscope, esophageal remnants and adherent adipose tissue were removed, and the tracheal sections were opened longitudinally, rinsed in PBS, and rotated in fresh DMEM containing 0.

The culture medium on the basolateral side of the filters was replaced every 48 hours. Apical medium was removed 4 days post seeding and then every time the basolateral culture medium was changed, and cells were cultured for additional days at an air-liquid interface.

Transepithelial resistance was monitored every other day when culture medium was replaced by use of an epithelial voltohmmeter WPI, Sarasota, FL. Organotypic cultures. AT2 cells were cultured as organoids as previously described Surface area of the organoids was measured by marking area across the colonies and then measured with the ImageJ software Figure S6. The fixed organoids were immunostained with AT1 and AT2 antibodies. Ussing chamber recordings. Measurements of short-circuit current Isc in MTE monolayers were performed as described previously 73 , Each solution was iso-osmolalic.

A mV pulse of 1-sec duration was imposed every 10 sec to monitor transepithelial resistance. Data were collected with the Acquire and Analyze program version 2. When the Isc level reached a plateau, drugs were pipetted to the apical compartment. The stoichiometry of native ENaC complexes is unclear to date.

Oocytes were surgically removed from appropriately anesthetized adult female Xenopus laevis Xenopus Express. Briefly, the ovarian tissue was removed from the frog under anesthesia by tricaine-S Western Chemicals through a small incision in the lower abdomen. Ovarian lobes were removed and digested in OR-2 calcium-free medium in mM: The two-electrode voltage-clamp technique was used to record whole-cell currents 48 h post injection.

Oocytes were impaled with two electrodes filled with 3M KCl, having resistance of 0. A TEVA voltage-clamp amplifier Dagan was used to clamp oocytes with concomitant recording of currents. Two reference electrodes were connected to the bath. The continuously perfused bathing solution was ND medium in mM: Whole-cell currents were recorded as previously reported Data were sampled at the rate of Hz and filtered at 50 Hz.

Germline transmitting mice were genotyped by Southern blotting and PCR for the transmission of the transgene. Surface biotinylation and immunoblotting assays. Quantified proteins were probed with specific monoclonal anti-HA antibody. Confocal imaging of organoids. Monolayers and organoids were scanned for Z sections with optimal width from top to bottom. Continuous images were stacked separately for pdpn and sftpc to count the number of positive cells for both by using plugin for cell counter in the Image J.

For three-dimensional reconstruction, a series of optical sections that were obtained by confocal microscope were collected at 0. FACS assays of differentiation and proliferation. Gates for both colours were set by unstained cells and isotype controls for each antibody.

Immunohistology of lung tissues and visualization of AT1 and AT2 cells. Mouse tissue sections were blocked using a proprietary blocking solution from a M. Nuclei were stained with Hoechst Thermo Fisher Scientific. Tissue staining images were taken by Zeiss LSM confocal systems Zeiss for immunofluorescence staining. After fixation, organoids were dehydrated with increasing ethanol grades and embedded as paraffin blocks.

EdU assay for DNA synthesis. Statistical analysis. Differences between means were tested for statistical significance using paired or unpaired t-tests or their non-parametric equivalents as appropriate. Differences between groups were judged using analysis of variance. We thank Dr. Dong-Ming Su from the University of North Texas for their kind technical support for transgenic mice.

All authors edited and reviewed the manuscript. S performed FACS. N performed ex vivo alveolar fluid clearance in human lungs. M and H. Europe PMC requires Javascript to function effectively. Recent Activity. The snippet could not be located in the article text. This may be because the snippet appears in a figure legend, contains special characters or spans different sections of the article.

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Corresponding author: H. Competing Interests: The authors have declared that no competing interest exists. Received May 27; Accepted Sep Associated Data Supplementary Materials Supplementary figures and tables. Abstract Lung epithelial sodium channel ENaC encoded by Scnn1 genes is essential for maintaining transepithelial salt and fluid homeostasis in the airway and the lung.

Keywords: alveolar type epithelial cells, epithelial sodium channels, self-renewal, humanized transgenic mouse line. Open in a separate window. Figure 1. Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Discussion The main findings of this study can be summarized as follows. Methods Human AT2 cell isolations. Supplementary Material Supplementary figures and tables. Click here for additional data file. Acknowledgments We thank Dr. Data availability The raw data that support the findings of this study are available from the corresponding authors upon request.

Author contributions All authors edited and reviewed the manuscript. References 1. J Biol Chem. A new channel type with homologies to Caenorhabditis elegans degenerins. FEBS Lett. Degenerin sites mediate proton activation of deltabetagamma-epithelial sodium channel. Chalfie M, Wolinsky E. The identification and suppression of inherited neurodegeneration in Caenorhabditis elegans.

Driscoll M, Chalfie M. The mec-4 gene is a member of a family of Caenorhabditis elegans genes that can mutate to induce neuronal degeneration. Epithelial sodium channel related to proteins involved in neurodegeneration [see comments] Nature. The epithelial sodium channel delta-subunit: new notes for an old song. Am J Physiol Renal Physiol.

Kellenberger S, Schild L. Physiol Rev. The contribution of epithelial sodium channels to alveolar function in health and disease. Annu Rev Physiol. Acute respiratory distress syndrome. Nat Rev Dis Primers. Role of epithelial sodium channels ENaC in the regulation of lung fluid homeostasis. Delta-subunit confers novel biophysical features to alpha beta gamma-human epithelial sodium channel ENaC via a physical interaction. The delta-subunit of the epithelial sodium channel ENaC enhances channel activity and alters proteolytic ENaC activation.

The human degenerin MDEG, an amiloride-sensitive neuronal cation channel, is localized on chromosome 17q Early death due to defective neonatal lung liquid clearance in alpha ENaC -deficient mice. Nat Genet. Hummler E, Planes C. Importance of ENaC-mediated sodium transport in alveolar fluid clearance using genetically-engineered mice. Cell Physiol Biochem.

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Wehrschetz-Sigl, A. Hasmann, G. Guebitz, A. Pol, Swati V. Pol, and Aharon Gedanken Europ. George, A. Shirly Ben-David Makhluf, I. Genish and A. Marciano, J. Nanoparticle Res. Shames, Ya. Mukovskii, E. Rapoport, A. Moshkovich, V. Gedanken, Yu. Koltypin, E. Sominski, G. Halperin, I. Etsion Wear , — Hai, A. Tsindlekht, I. Felner, E. Mukovskii, C. On Magnet. Richman, S. Willick, N.

Skirtenko, A. Cluster Science a special issue dedicated to the 75 Birthday of Prof. Rao 20, Irzh, I. Genish, ; L. Chen, Y-C. Ling, L. Klein, A. Perkas, Sh. Magdassi, T. Zioni, M. Royz, Z. Maor, A. Abu-Much, A. Chem C. Polymer Science A: Polymer Chem. George, S. Pol, J. Chen, A. Gedanken, N. Perkas and Z. Shames, M. Auslender, E. Mukovskii , J. Luvchik, J. Calderon-Moreno, N. Veglio, J. Tamarit Advanced Materials , 20 , Gedanken, Microp.

Haas, A. Rozenberg, Ya. Gedanken, Journal of Magnetism and Magnetic Materials , e8—e11 Vilas G. Pol, Aharon Gedanken, Langmuir 24, Liu, L. Chen, H. Kai, S. Zhou, U. Frydman, A. Luvchik, A. Gedanken, Europ. Rich and A. Gedanken, Adv. Shames, D. Mogilyansky, I. Sominskii, A. Mukovskii, Phys. B 78 , Jacob, Somashekarappa Mallenahalli, Leonid A.

Porphyrines and Phthalocyanines. Chem C , Jacob, A. George, V. Gabashivili, M. Cai, A. Feng, M. Ruthkosky, J. Borodianskiy, V. Mazurovsky, M. Zinigrad, A. Gedanken Isr. Shamugam, A. Mazurovsky, A. Gedanken, M. Zinigrad, Mater, Sci. Kalai Selvan, C. Augustin, V. Sepelak, L. John Berchmans, C. Sanjeeviraja, A. Zitoun and A. Gedanken, Nanotechnology 19 , Rozenberg, ; M.

Auslender, ; A. Shames, ; Ya. Mukovskii, ; E. Sominski, E. Gedanken J. Metals , Auslender, I. Pestun, Ya. Pol ; Swati V. Ultrasonics Sonochemistry, S. Avivi Levi and A. Gedanken 14 , , Rozenberg, G. Gorodetsky, I. Pang and A. Gedanken, Current Applied Physics, , 7, Patra, Ayelet Odani, Vilas G. Pol, and A. Nemet, Ehud I.

Pol, V. Grinblat, R. Kalai Selvan, V. Kessler, G. Spijksma, G. Seisenbaeva, and S. Gohil, J. Amiryan, S. Dubinsky, S. Gazit, A. Polymer Science , Gabashvili, Mater. Gedanken, Ultrasonic Sonochem. Rozenberg, S. Banerjee, I. Sominski and A. Pol, Vilas G. Ben-Ishay, A. Kotlyar, N. Amiryan, M. Meyer, W. Langmuir 23 , Gedanken, and Ya. Tuval and A. Pol, Aharon Gedanken, I. Felner, I.

Sung, ; S. Asai, Inorg. Jung, , A. Koltypin, Phys. Koltypin, V. Gedanken, D. Aurbach J. Irzh, N. Gedanken, Langmuir 23 , Gabashvili, Dana. B , Pol and A. Jung, Ya. Rozenberg,1, A. Auslender, G. Banerjee, D. B 76 , Solovyov, Evangelia Xenogiannopoulou,. Perkas, V. Gorodetsky, E. Gedanken and Ya.

Kessler, and Aharon Gedanken, New J. Abu-Much, U. Jacob, Jimmy C. Yu, Aharon Gedanken, Chem. Pang, G. Chem B , Jung, M. Mukovskii, Submitted to J. Augustin, C. Sanjeeviraja, V. Zhong, F. Chen, T. Ang, Y. Han, W. Lim, A. Gedanken, Inorg. B ; , Kim, D. Glass Phys. Landau, Aharon Gedanken, Langmuir 22, Pol, P.

George, B. Markovsky and A. Kim, A. Tver'yanovich, D. Lee, and B. Pol, Aharon Gedanken Carbon 44 , Turgeman and A. Gedanken, Cryst. Zhou, K. Huang, U. Frydrnan, A. Langmuir 22 , Pol, Jose M. Calderon Moreno, Aharon Gedanken, Carbon 44 , Rangari, D.

Srivastava, A. Grzanka, and W. Yatsunenko, V. Osinniy and T. Story, Appl. Sivakumar, A. Bhattacharya, Y. Yeshurun, W. Zhong, Y. Du, I. Felner, J. Turgeman, O. Gershevitz, O. Palchik, M. Deutsch, B. Ocko, A. Gedanken, C. Gofer, J. Edited by C. Hoz, L. Luong, Aharon Gedanken, J. Oliete , T. Seisenbaeva, M. Sung, S. Asai, and A. B, , Pol, Gulaim Seisenbaeva, Vadim G. Kessler, and A. Bahadur, S. Vitta, O. Prakash, Narosa Publishing House, , Pages.

Nikitenko, Yu. Koltypin, I. Yeshurun, A. Shames, J. Jiang, V. Markovich, G. Gorodetsky, A. Rana, V. Pol, I. Frydman, Adv. Gedanken, Ultrasonic Sonochemistry 11 , - , Gedanken, V. Gorodetsky, G. Pang, and A. Gedanken, B. Atkas et al. Brukental, Y.

Nowik, Chem. Qiu, Vilas G. Calderon-Moreno, and Aharon Gedanken, Langmuir 20 , Kumar, J. Gnanaraj, G. Salitra, A. Abramov, A. Aurbach, J. Soupart and J. Rousche, Journal of Solid State Electrochemistry 8, - Koltypin, N. Perkas, M. Besson, L. Vradman, M. Hershkovitz, M. Landau, Trans. Tang, W. Huang Huang, S. Liu, Y. Wang, O. Palchik, Yu. Koltypin, D. Pickup, E. Jortner 70th birthday. State Chem. Nanoparticles Research 6, Srivastava, N. Seisenbaeva, Y. Kessler and A.

Van-Eck, A. Gedanken, Ultrason. Ganesh Kumar, D. Aurbach, and A. Gedanken Ultrasonic Sonochemistry 10, 17 Landau, M. Herskowitz, V. Ezersky, M. Talianker, S. Gedanken, and J. Calderon-Moreno, Chem. Pol, M. Motiei, A. Mastai, Chem. Topaz, A. Rubowitz, E. Assia, M.

Rosner, D. Meyerstein, N. Meyerstein and A. Perkas, H. Buchkremer, N. Rana, X. Xu, Y. Yeshurun and A. Palchik, A. Odani, A. Nimberger, B. Markovsky, E. Levi, V. Kumar, M. Gedanken, P. Dan, D. Aurbach, Journal of Power Sources, , Menzler, D. Lavergnat, F. Tietz, E. Djurado, W. Fischer, G. Pang, A. Gedanken, H. New J. Gedanken, Stud. Palchik, V. Pol, E. Sominska, Yu.

Perkas, O. Gedanken, Submitted to J. Gnanaraj, V. Gedanken and D. Aurbach, Electrochem. Gnanaraj, S. Ben-David, D. Aurbach, Chem. Gabashvili, G. Seisenbaeva, V. Kessler, L. Zhang, J. Yu, A. Gedanken, Current Science, 85, No. Avivi Levi, , Y.

Dror and A. Topaz, E. Assia, D. Meyerstein, A. Gedanken, Invest. Assia, A. Rubowitz, M. Topaz, Invest. Palchik, O. Koltypin, Y. Diamant, Y. Sominska, E. Grisaru, O. Slifkin, A. Hong-liang Li, Ying-chun Zhu, O. Gedanken, I. Chem B , - Jeevanandam, Yu.

B 90, Koltypin, S. Nikitenko, A. Pol, D. Srivastava, O. Gedanken, Langmuir 18, Wolfus, A. Shaulov, Y. Yeshurun, I. Novik, Yu. Rana, A. Yeshurun, X. Xu, J. Weiss, J. Rozenberg G. Arsenov, D. Shulyatev, Ya. Mukovskii, A.

Gedanken, G. Pang, J. Harpeness, O. Palchik, S. Amiel, M. Weiss, Chem. Motiei, J. Calderon-Moreno, A. Socol, O. Abramson, A. Meshorer, A. Aurbach, A. Koltypin, O. Palchik, and A. Gedanken, Biochemical Journal , Topaz, M. Meyerstein, Ultrasound in Medicine and Biology 28 , Huang, X. Tang, I. Felner, Yu. Kumar Rana, Y. Gedanken Adv. Pol, R. Reisfeld, and A. Pol, O.

Felner, Appl. Nimberger, E. Levi, E. Gedanken Chem. Stanislav Kishinevski, Sergei I. Pickup, Ernst R. Publications in Wang, S. Chen, X. Tang, O. Zaban, Yu. Vijaya Kumar, R. Elgamiel, Y. Gedanken , Langmuir, 17 , Palchik, R. Kerner, A. Weiss, M. Slifkin, V. Palchik J. Gadanken J.

Vijaya Kumar, Y. Mastai, Y. Diamant, A. Kerner, O. Pang, E. Sominska, H. Colfen, Y. Mastai, S. Avivi, Yu, Koltypin, A. Gedanken, Nanoletters 1, Liu, W. C Dose-response curve for amiloride. Current levels at each dose were plotted against applied dose of amiloride and then fitted raw data with the Hill equation to calculate IC 50 value 1. Dashed line red is generated with the fitted parameters. All four ENaC subunits are co-expressed in many tissues 29 , 36 , Scale bars at the left bottom corner are for time and current amplitude.

The current traces were digitized at the membrane potential of mV. On the other hand, aprotinin did not alter the current amplitude significantly. C Identification of subunit responsible for plasmin activation. Control, without pre-treatment. NI, non-injected oocytes as negative controls for ENaC expression. Cells treated with the same volume of saline - were controls for serine proteases. Inducible expression of human Scnn1d in mouse airway and lung epithelium. In contrast to human and other species, mice do not express Scnn1d gene 8 , In addition, the band at kDa was only found in polarized MTE cells.

A Schematic design for developing humanized mouse strain. Two lox and one stop codon were placed just prior to this construct. C Transcripts of ENaC subunits in the lung. Bottom panels are for alveolar type 2 AT2 cells stained with sftpc as a biomarker. F Immunoblotting biotinylated apical proteins recognized by anti-HA antibody.

These blots represent three experiments with similar results. Primary MTE cells were cultured at the air-liquid interface for days. Arrows indicate the time to add designed concentrations of amiloride. Raw data were fitted with the logistic function to compute k i values for amiloride. The resulting k i values were 0. D Dose-response curve for amiloride in mAT2 cells. Resistance was read with a chopstick meter when the culture medium was replaced every other day. ENaC is expressed in pulmonary epithelial stem and progenitor cells and implicated in the repair of injured epithelial and other tissues 40 - Scale bar, 1 mm.

B Organoid number per well. E AT2 renewal and differentiation into AT1 cells. Polarized mAT2 monolayers on day 7 post seeding were stained with EdU green. Student t-test. J Total surface area of organoids per well. The main findings of this study can be summarized as follows. We established a novel humanized transgenic mouse model conditionally expressing full-length human Scnn1d gene.

We found that the Scnn1d gene is a regulator of alveolar progenitor AT2 cells and also functions as a critical transepithelial sodium transport pathway. The model of ion selectivity filter is composed of three layers vertically 45 , The potential mechanisms are related to the augmented proliferation of alveolar progenitor mouse AT2 cells. On the other hand, we cannot exclude the potential contribution of Scnn1a , Scnn1b , and Scnn1g to the fate of AT2 cells.

Broadly speaking, the linage of AT2 cells could be regulated by more than one gene. To the best of our knowledge, this is the first study on the regulation of AT2 fate by Scnn1d genes. These could be due to considerable loss of AT2 cells during isolation and the difference in niche for AT2 cells between in vivo and in vitro.

The scenario for alveologenesis in fetal lungs and regeneration of the air sacs in injured lungs could fit the in vitro 3D cultures. Technically, this discrepancy could be resulted from the unidentical affinity between pdpn and ICAM antibodies.

We could not use the same antibody, either pdpn or ICAM, for both confocal imaging and flow cytometry for their diverse applications. Whether the contribution of delta-ENaC to alveolar fluid clearance is mediated by its proliferative effects on AT2 cells is unclear. This well-controlled strategy provide rigorous data. The clinical implications of Scnn1d in airway fluid homeostasis is emerging.

They acquired P. In addition to the function of Scnn1d in human nasal epithelium 22 , we for the first time confirmed the contribution of Scnn1d gene to alveolar fluid clearance. Human AT2 cell isolations. Human AT2 cells were isolated with a modified protocol 66 , Briefly, distal lung tissue was obtained and dissected into rough 5 cm 3 pieces.

Tissue was compressed with autoclaved Kim wipes to remove as much liquid as possible and further dissected into 1 cm 3 pieces. Fungizone was added for the final 30 minutes of the digest. Finally, red blood cells were removed using the red blood cell lysis buffer Sigma-Aldrich. Alveolar fluid clearance in human lungs ex vivo.

All patients were given oral and written informed consent forms. Human lung tissues were obtained during pulmonary resection surgeries with lung cancer at the First Affiliated Hospital of CMU. There were no fibrous or emphysematous lesions as assessed by preoperative chest CT scanning. Human lung segments were prepared, and AFC measurements were done as described previously Briefly, the segmental bronchus was occluded by a Fr.

Alveolar fluid was aspirated 60 minutes after instillation. In vivo alveolar fluid clearance AFC in mice. In vivo AFC rate was measured as previously described 68 - To maximize the collection of instilled BSA solution, the diaphragm was dissected. Methodological concerns were addressed by using amiloride to inhibit water movement to confirm that measurements accurately reflected AFC.

The endogenous murine albumin would lead to an overestimation of AFC rates. This potential bias was corrected by measuring murine albumin AssayPro, St. Final AFC values were corrected removing murine albumin present in the aspirate. In addition, we used an aliquot of the instillate that had been delivered into the lungs and removed within 5 min, rather than freshly prepared naive instillate per se. Any pre-existing edema or leaking murine plasma proteins would be excluded for it dilutes both the control at time point 0 min P i and final samples P f.

Mouse AT2 cell isolation and monolayer cultures. Mouse tracheal epithelial cell isolations and monolayer cultures. Briefly, mice were anesthetized with intraperitoneal injection of ketamine HCl and xylazine. The trachea proximal to the bronchial bifurcation was isolated and removed. The resected section was immediately placed in PBS. Under a dissecting microscope, esophageal remnants and adherent adipose tissue were removed, and the tracheal sections were opened longitudinally, rinsed in PBS, and rotated in fresh DMEM containing 0.

The culture medium on the basolateral side of the filters was replaced every 48 hours. Apical medium was removed 4 days post seeding and then every time the basolateral culture medium was changed, and cells were cultured for additional days at an air-liquid interface.

Transepithelial resistance was monitored every other day when culture medium was replaced by use of an epithelial voltohmmeter WPI, Sarasota, FL. Organotypic cultures. AT2 cells were cultured as organoids as previously described Surface area of the organoids was measured by marking area across the colonies and then measured with the ImageJ software Figure S6.

The fixed organoids were immunostained with AT1 and AT2 antibodies. Ussing chamber recordings. Measurements of short-circuit current Isc in MTE monolayers were performed as described previously 73 , Each solution was iso-osmolalic. A mV pulse of 1-sec duration was imposed every 10 sec to monitor transepithelial resistance. Data were collected with the Acquire and Analyze program version 2. When the Isc level reached a plateau, drugs were pipetted to the apical compartment.

The stoichiometry of native ENaC complexes is unclear to date. Oocytes were surgically removed from appropriately anesthetized adult female Xenopus laevis Xenopus Express. Briefly, the ovarian tissue was removed from the frog under anesthesia by tricaine-S Western Chemicals through a small incision in the lower abdomen. Ovarian lobes were removed and digested in OR-2 calcium-free medium in mM: The two-electrode voltage-clamp technique was used to record whole-cell currents 48 h post injection.

Oocytes were impaled with two electrodes filled with 3M KCl, having resistance of 0. A TEVA voltage-clamp amplifier Dagan was used to clamp oocytes with concomitant recording of currents. Two reference electrodes were connected to the bath. The continuously perfused bathing solution was ND medium in mM: Whole-cell currents were recorded as previously reported Data were sampled at the rate of Hz and filtered at 50 Hz.

Germline transmitting mice were genotyped by Southern blotting and PCR for the transmission of the transgene. Surface biotinylation and immunoblotting assays. Quantified proteins were probed with specific monoclonal anti-HA antibody. Confocal imaging of organoids. Monolayers and organoids were scanned for Z sections with optimal width from top to bottom. Continuous images were stacked separately for pdpn and sftpc to count the number of positive cells for both by using plugin for cell counter in the Image J.

For three-dimensional reconstruction, a series of optical sections that were obtained by confocal microscope were collected at 0. FACS assays of differentiation and proliferation. Gates for both colours were set by unstained cells and isotype controls for each antibody.

Immunohistology of lung tissues and visualization of AT1 and AT2 cells. Mouse tissue sections were blocked using a proprietary blocking solution from a M. Nuclei were stained with Hoechst Thermo Fisher Scientific. Tissue staining images were taken by Zeiss LSM confocal systems Zeiss for immunofluorescence staining.

After fixation, organoids were dehydrated with increasing ethanol grades and embedded as paraffin blocks. EdU assay for DNA synthesis. Statistical analysis. Differences between means were tested for statistical significance using paired or unpaired t-tests or their non-parametric equivalents as appropriate. Differences between groups were judged using analysis of variance.

We thank Dr. Dong-Ming Su from the University of North Texas for their kind technical support for transgenic mice. All authors edited and reviewed the manuscript. S performed FACS. N performed ex vivo alveolar fluid clearance in human lungs. M and H. Europe PMC requires Javascript to function effectively. Recent Activity. The snippet could not be located in the article text.

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