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20 results on '"Gatti, A. P."'

1. Global wealth disparities drive adherence to COVID-safe pathways in head and neck cancer surgery

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Schache, Andrew G, Shaw, Richard, Ho, Michael Wing Sung, Winter, Stuart C, Glasbey, James, Ganly, Ian, Batstone, Martin, Biel, Juan Rey, Nankivell, Paul C, Simon, Christian, Omar, Omar, Simoes, Joana F F, Nepogodiev, Dmitri, Bhangu, Aneel, Pinkney, Tom, McGill, Laura, Perry, Rita, Hughes, Terry, Jackson, Richard, Siaw-Acheampong, Kwabena, Benson, Ruth A, Bywater, Edward, Chaudhry, Daoud, Dawson, Brett E, Evans, Jonathan P, Glasbey, James C, Gujjuri, Rohan R, Heritage, Emily, Jones, Conor S, Kamarajah, Sivesh K, Khatri, Chetan, Khaw, Rachel A, Keatley, James M, Knight, Andrew, Lawday, Samuel, Li, Elizabeth, Mann, Harvinder S, Marson, Ella J, McLean, Kenneth A, Mckay, Siobhan C, Mills, Emily C, Pellino, Gianluca, Picciochi, Maria, Taylor, Elliott H, Tiwari, Abhinav, Trout, Isobel M, Venn, Mary L, Wilkin, Richard J W, Ho, Michael W S, Nankivell, Paul, Abbott, Tom E F, Adamina, Michel, Ademuyiwa, Adesoji O, Agarwal, Arnav, Alameer, Ehab, Alderson, Derek, Alakaloko, Felix, Albertsmeiers, Markus, Alser, Osaid, Alshaar, Muhammad, Alshryda, Sattar, Arnaud, Alexis P, Augestad, Knut Magne, Ayasra, Faris, Azevedo, José, Bankhead-Kendall, Brittany K, Barlow, Emma, Blanco-Colino, Ruth, Brar, Amanpreet, Minaya-Bravo, Ana, Breen, Kerry A, Bretherton, Chris, Buarque, Igor Lima, Burke, Joshua, Caruana, Edward J, Chaar, Mohammad, Chakrabortee, Sohini, Christensen, Peter, Cox, Daniel, Cukier, Moises, Cunha, Miguel F, Davidson, Giana H, Desai, Anant, Saverio, Salomone Di, Drake, Thomas M, Edwards, John G, Elhadi, Muhammed, Emile, Sameh, Farik, Shebani, Fiore, Marco, Edward Fitzgerald, J, Ford, Samuel, Garmanova, Tatiana, Gallo, Gaetano, Ghosh, Dhruv, Gomes, Gustavo Mendonça Ataíde, Grecinos, Gustavo, Griffiths, Ewen A, Gründl, Madalegna, Halkias, Constantine, Harrison, Ewen M, Hisham, Intisar, Hutchinson, Peter J, Hwang, Shelley, Isik, Arda, Jenkinson, Michael D, Jonker, Pascal, Kaafarani, Haytham M A, Kolias, Angelos, Kruijff, Schelto, Lawani, Ismail, Lederhuber, Hans, Leventoglu, Sezai, Litvin, Andrey, Loehrer, Andrew, Löffler, Markus W, Lorena, Maria Aguilera, Madolo, Maria Marta, Major, Piotr, Martin, Janet, Mashbari, Hassan N, Mazingi, Dennis, Metallidis, Symeon, Mohan, Helen M, Moore, Rachel, Moszkowicz, David, Moug, Susan, Ng-Kamstra, Joshua S, Maimbo, Mayaba, Niquen, Milagros, Ntirenganya, Faustin, Olivos, Maricarmen, Oussama, Kacimi, Outani, Oumaima, Parreno-Sacdalanm, Marie Dione, Pata, Francesco, Rivera, Carlos Jose Perez, Pinkney, Thomas D, van der Plas, Willemijn, Pockney, Peter, Qureshi, Ahmad, Radenkovic, Dejan, Medina, Antonio Ramos-De la, Roberts, Keith, Roslani, April C, Rutegård, Martin, Santos, Irène, Satoi, Sohei, Sayyed, Raza, Schache, Andrew, Schnitzbauer, Andreas A, Seyi-Olajide, Justina O, Sharma, Neil, Shu, Sebastian, Soreide, Kjetil, Spinelli, Antonino, Stewart, Grant D, Sund, Malin, Sundar, Sudha, Tabiri, Stephen, Townend, Philip, Tsoulfas, Georgios, van Ramshorst, Gabrielle H, Vidya, Raghavan, Vimalachandran, Dale, Warren, Oliver J, Wedderburn, Duane, Wright, Naomi, Boccalatte, L A, Batstone, M, Hodge, R, Abeloos, J, De Backer, T, De Ceulaer, J, Dick, C, Diez-Fraile, A, Lamoral, P, Spaas, C, Schrijvers, D L A L, Willemse, E B M, Faris, C, Maariën, S, Van Haesendonck, G, Van Laer, C, Deron, P, Abdallah, E A, Carvalho, G B, Kowalski, L, Vartanian, J, Gatti, A P, Nardi, C N, Oliva, R N L, Salem, M C, Cheng, D, MacNeil, D, Martin, J, Mayer, R, Groot, G, Acosta, L, Mejia, M, Perez, C J, Lorencin, M, Luksic, I, Mamic, M, Ashoush, F M, Osman, N A, Safwat Shahine, M, Eldaly, A, Elfiky, M M A, Amin, A, Elmorsi, R, Refky, B, Essa, M M, Mengistu, G Mengesha, Dakpé, S, Boucher, S, Ballouhey, Q, Laloze, J, Usseglio, J, Hoffmann, C, Gregoire, V, Lallemant, B, Blaurock, M, Reim, D, Boehm, A, Guntinas-Lichius, O, Hölzle, F, Modabber, A, Winnand, P, Kleeff, J, Lorenz, K, Ronellenfitsch, U, Schneider, R, Betz, C S, Böttcher, A, Busch, C, Möckelmann, N, Inhestern, J M, Greve, J, Hoffmann, T K, Laban, S, Vahl, J M, Agyeman-Prempeh, A, Aning, D, Barnor, I, Darko-Asante, R, Dzogbefia, M, Gaveh, V, Gyimah, D, Issahalq, M D, Konney, A, Poku, M, Adjeso, T, Akornor, E T, Amankwaa, W O, Antwi, D A, Apppiah-Thompson, P, Damah, M, Kumi, E O, Manan, L, Murphy, J P, Osei, L, Setuagbe, J, Arkadopoulos, N, Danias, N, Economopoulou, P, Frountzas, M, Kokoropoulos, P, Larentzakis, A, Michalopoulos, N V, Nastos, K, Parasyris, S, Pikoulis, E, Selmani, J, Sidiropoulos, T A, Vassiliu, P, Kalfountzos, C E, Chatziioannou, I, Corais, C, Gkrinia, E, Ntziovara, A, Saratziotis, A, Antoniadis, K, Orestis, O, Tatsis, D, Baili, E, Charalabopoulos, A, Liakakos, T, Schizas, D, Spartalis, E, Syllaios, A, Zografos, C, Aguilera-Arévalo, M, Misra, S, Pareek, P, Vishnoi, J, Chappity, P, Kar, M, Muduly, D K, Sultania, M, Agarwal, S, Garg, P K, Maharaj, D D, Majumdar, K S, Mishra, N, Poonia, D, Seenivasagam, R K, Singh, M P, Tiwari, A R, Penumadu, P, Rajan, S, Kumar, S, Raychowdhury, R, Ghodke, R, Barry, C, Callanan, D, Dias, A, Haung, L, Ionescu, A, Sheahan, P, Lennon, P, Fitzgerald, C, Mizrachi, A, Deganello, A, Pellini, R, Pichi, B, Lemma, F, Marino, M V, Bergonzani, M, Varazzani, A, Bussu, F, Perra, T, Piras, A, Porcu, A, Rizzo, D, Campisi, G, Cordova, A, Franza, M, Rinaldi, G, Toia, F, Gianni, A, Giannini, L, Gordini, L, Baldini, E, Conti, L, De Virgilio, A, Ferreli, F, Gaino, F, Mercante, G, Spriano, G, Ansarin, M, Chu, F, De Berardinis, R, Ietrobon, G, Tagliabue, M, Ionna, F, Baietti, A, Maremonti, P, Neri, F, Prucher, G, Ricci, S, Casaril, A, Nama, M, Cotoia, A, Lizzi, V, Vovola, F, Bruzzaniti, P, Familiari, P, Lapolla, P, Marruzzo, G, Mingoli, A, Ribuffo, D, Cipriani, R, Contedini, F, Lauretta, M, Marchetti, C, Melotti, M, Pignatti, M, Pinto, V, Pizzigallo, A, Ricotta, F, Tarsitano, A, Catarzi, L, Consorti, G, Abdulwahed, E A, Alshareea, E A, Biala, M I, Ghmagh, R J, Ibrahim, A F, Liew, Y T, Alvarez, M R, Arrangoiz, R, Cordera, F, Gómez-Pedraza, A, Soulé-Martínez, C E, Becerril, O S, Becerra, GFC, Arkha, Y, Bechri, H, El Ouahabi, A, Oudrhiri, M, Benkabbou, A, Majbar, M, Mohsine, R, Souadka, A, Lageju, N, Schreuder, W H, Hardillo, J, de Bree, R, Schweitzer, D, Adeyeye, A A, Enoch, E E, Sholadoye, T S T T, Wuraola, F, Oyelakin, O, Khokhar, M I, Ayub, B, Walędziak, M, Szewczyk, M, Faria, C, Cardoso, P, Castro Silva, J, AlKharashi, E, Jelovac, D, Petrovic, M, Sumrak, S, Asceric, R R, Bojicic, J M, Kovacevic, B M, Krdzic, I D, Milentijevic, M A, Milutinovic, V Z, Stefanovic, Z B, Villacampa, J M, Jiménez Carneros, V, Salazar Carrasco, A, Carabias Hernandez, A, Alonso Lamberti, L, León Ledesma, R, Jiménez Miramón, F J, Jover Navalón, J M, Garcia Quijada, J, Ramos Rodriguez, J, Valle Rubio, A, Vilaseca, I, Escartin, J, Estaire-Gomez, M, Padilla Valverde, D, Tousidonis, M, Lopez, F, Deandrés-Olabarria, U, Durán-Ballesteros, M, Fernández-Pablos, F, Ibáñez-Aguirre, F, Sanz-Larrainzar, A, Ugarte-Sierra, B, Di Martino, M, Prada, J, Jariod-Ferrer, U M, Landaluce Olavarria, A, Rey-Biel, J, Díaz de Cerio, P, Sánchez Barrueco, A, Lindqvist, E K, Sund, M, Piantanida, R, Giger, R, Hool, S, Müller, S A, Stoeckli, S J, Simon, C, Toutounji, T, Al assaf, A, Hammed, A M, Hammed, S M, Mahfoud, M, Arikan, A, Yalkin, Ö, İflazoğlu, N, Isik, A, Leventoglu, S, Aydemir, L, Basaran, B, Sen, C, Comert -Ulusan, M, Saracoglu, K T, Saracoglu, A, Mantoglu, B, Kucuk, G, Aygun, N, Baran, E, Tanal, M, Eray Tufan, A, Uludag, M, Gürkan Yetkin, S, Yigit, B, Calik, B, Demirli Atici, S, Kaya, T, Sikakulya, F K, Abdel-Galil, K M A H, Lowe, T, Durrani, A J, Habeeb, A, Irune, E, Luke, L, Masterson, L, Murphy, S H, Segaren, N, Walker, C, Waseem, S, Jones, T M, Loh, C, Pringle, S, Schache, A G, Shaw, R J, Stenhouse, J, Armstrong, M, Sood, S, Sutton, D, Thomas, S, Clarke, P, Winter, S C, Hislop, S, Counter, P R, Ghazali, N, Lloyd, C, Prabhu, V, Godden, D, Whitley, S, Butler, C, Nash, R, El-Boghdadly, K, Fry, A, Niziol, R, De, M, Gill, C K, Crank, S, Mace, A D, Ho, M, Mair, M, Kothari, P, Homer, J, Sainuddin, S, Egan, R J, Kittur, M, Burgess, C, O'Hara, J, Manickavasagam, J, McDonald, C, Burrows, S, Java, K R, Katre, C, Ahmed, A, Siddique, H, King, E, Ramchandani, P, Naredla, P R, Brennan, P, Ringrose, T, Schmidt, F, Mak, J K C, Nankivell, P, Parmar, S, Sharma, N, Douglas, C, McCaul, J, Dhanda, J, Kyzas, P, Vassiliou, L, Kumar, A, Husband, A, Hulbert, J, Ingrams, D, Parkin, R, Varley, I, Gahir, D, George, A, Zakai, D, Bater, M, Surwald, C, Devlin, B, Leonard, C G, Pigadas, N, Snee, D, Singh, R P, Hyde, N C, Paley, M, Cocks, H, Wilson, A, Choi, D, Kerawala, C J, Riva, F, Dickason, A, Semple, C J, Schilling, C, Walton, G, Rees-Stoner, O, Scott, N, Nixon, I J, Tighe, D, Mattine, S, Chu, M M H, Pothula, V, Lee, W, Brown, L, Ganly, I, Alpert, N, Illezeau, C N, Miles, B, Rapp, J, Taioli, E, Azam, M T, Choudhry, A J, Marx, W, Stein, J, Ying, Y, Gross, N D, Almasri, M, Joshi, R, Kulkarni, G, Marwan, H, Mehdi, M, Sumer, B, Department of Surgery, HUS Abdominal Center, Schache, Andrew G., Shaw, Richard, Omar, Omar, Emile, Sameh, Farik, Shebani, Fiore, Marco, Edward Fitzgerald, J., Ford, Samuel, Garmanova, Tatiana, Gallo, Gaetano, Ghosh, Dhruv, Gomes, Gustavo Mendonça Ataíde, Grecinos, Gustavo, Simoes, Joana F. F., Griffiths, Ewen A., Gründl, Madalegna, Halkias, Constantine, Harrison, Ewen M., Hisham, Intisar, Hutchinson, Peter J., Hwang, Shelley, Isik, Arda, Jenkinson, Michael D., Jonker, Pascal, Nepogodiev, Dmitri, Kaafarani, Haytham M. A., Kolias, Angelos, Kruijff, Schelto, Lawani, Ismail, Lederhuber, Hans, Leventoglu, Sezai, Litvin, Andrey, Loehrer, Andrew, Löffler, Markus W., Lorena, Maria Aguilera, Bhangu, Aneel, Madolo, Maria Marta, Major, Piotr, Martin, Janet, Mashbari, Hassan N., Mazingi, Dennis, Metallidis, Symeon, Minaya-Bravo, Ana, Mohan, Helen M., Moore, Rachel, Moszkowicz, David, Pinkney, Tom, Moug, Susan, Ng-Kamstra, Joshua S., Maimbo, Mayaba, Niquen, Milagros, Ntirenganya, Faustin, Olivos, Maricarmen, Oussama, Kacimi, Outani, Oumaima, Parreno-Sacdalanm, Marie Dione, Pata, Francesco, McGill, Laura, Rivera, Carlos Jose Perez, Pinkney, Thomas D., van der Plas, Willemijn, Pockney, Peter, Qureshi, Ahmad, Radenkovic, Dejan, Medina, Antonio Ramos-De la, Roberts, Keith, Roslani, April C., Rutegård, Martin, Perry, Rita, Santos, Irène, Satoi, Sohei, Sayyed, Raza, Schache, Andrew, Schnitzbauer, Andreas A., Seyi-Olajide, Justina O., Sharma, Neil, Shu, Sebastian, Soreide, Kjetil, Hughes, Terry, Spinelli, Antonino, Stewart, Grant D., Sund, Malin, Sundar, Sudha, Tabiri, Stephen, Townend, Philip, Tsoulfas, Georgios, van Ramshorst, Gabrielle H., Vidya, Raghavan, Vimalachandran, Dale, Jackson, Richard, Warren, Oliver J., Wedderburn, Duane, Wright, Naomi, Boccalatte, L. A., Batstone, M., Hodge, R., Abeloos, J., De Backer, T., De Ceulaer, J., Dick, C., Siaw-Acheampong, Kwabena, Diez-Fraile, A., Lamoral, P., Spaas, C., Schrijvers, D. L. A. L., Willemse, E. B. M., Faris, C., Maariën, S., Van Haesendonck, G., Van Laer, C., Deron, P., Ho, Michael Wing Sung, Benson, Ruth A., Abdallah, E. A., Carvalho, G. B., Kowalski, L., Vartanian, J., Gatti, A. P., Nardi, C. N., Oliva, R. N. L., Salem, M. C., Cheng, D., MacNeil, D., Bywater, Edward, Martin, J., Mayer, R., Groot, G., Acosta, L., Mejia, M., Perez, C. J., Lorencin, M., Luksic, I., Mamic, M., Ashoush, F. M., Chaudhry, Daoud, Osman, N. A., Safwat Shahine, M., Eldaly, A., Elfiky, M. M. A., Amin, A., Elmorsi, R., Refky, B., Essa, M. M., Mengistu, G. Mengesha, Dakpé, S., Dawson, Brett E., Boucher, S., Ballouhey, Q., Laloze, J., Usseglio, J., Hoffmann, C., Gregoire, V., Lallemant, B., Blaurock, M., Reim, D., Boehm, A., Evans, Jonathan P., Guntinas-Lichius, O., Hölzle, F., Modabber, A., Winnand, P., Kleeff, J., Lorenz, K., Ronellenfitsch, U., Schneider, R., Betz, C. S., Böttcher, A., Glasbey, James C., Busch, C., Möckelmann, N., Inhestern, J. M., Greve, J., Hoffmann, T. K., Laban, S., Vahl, J. M., Agyeman-Prempeh, A., Aning, D., Barnor, I., Gujjuri, Rohan R., Darko-Asante, R., Dzogbefia, M., Gaveh, V., Gyimah, D., Issahalq, M. D., Konney, A., Poku, M., Adjeso, T., Akornor, E. T., Amankwaa, W. O., Heritage, Emily, Antwi, D. A., Apppiah-Thompson, P., Damah, M., Kumi, E. O., Manan, L., Murphy, J. P., Osei, L., Setuagbe, J., Arkadopoulos, N., Danias, N., Jones, Conor S., Economopoulou, P., Frountzas, M., Kokoropoulos, P., Larentzakis, A., Michalopoulos, N. V., Nastos, K., Parasyris, S., Pikoulis, E., Selmani, J., Sidiropoulos, T. A., Kamarajah, Sivesh K., Vassiliu, P., Kalfountzos, C. E., Chatziioannou, I., Corais, C., Gkrinia, E., Ntziovara, A., Saratziotis, A., Antoniadis, K., Orestis, O., Tatsis, D., Winter, Stuart C., Khatri, Chetan, Baili, E., Charalabopoulos, A., Liakakos, T., Schizas, D., Spartalis, E., Syllaios, A., Zografos, C., Aguilera-Arévalo, M., Misra, S., Pareek, P., Khaw, Rachel A., Vishnoi, J., Chappity, P., Kar, M., Muduly, D. K., Sultania, M., Agarwal, S., Garg, P. K., Maharaj, D. D., Majumdar, K. S., Mishra, N., Keatley, James M., Poonia, D., Seenivasagam, R. K., Singh, M. P., Tiwari, A. R., Penumadu, P., Rajan, S., Kumar, S., Raychowdhury, R., Ghodke, R., Knight, Andrew, Barry, C., Callanan, D., Dias, A., Haung, L., Ionescu, A., Sheahan, P., Lennon, P., Fitzgerald, C., Mizrachi, A., Deganello, A., Lawday, Samuel, Pellini, R., Pichi, B., Lemma, F., Marino, M. V., Bergonzani, M., Varazzani, A., Bussu, F., Perra, T., Piras, A., Porcu, A., Li, Elizabeth, Rizzo, D., Campisi, G., Cordova, A., Franza, M., Rinaldi, G., Toia, F., Gianni, A., Giannini, L., Gordini, L., Baldini, E., Mann, Harvinder S., Conti, L., De Virgilio, A., Ferreli, F., Gaino, F., Mercante, G., Spriano, G., Ansarin, M., Chu, F., De Berardinis, R., Ietrobon, G., Marson, Ella J., Tagliabue, M., Ionna, F., Baietti, A., Maremonti, P., Neri, F., Prucher, G., Ricci, S., Casaril, A., Nama, M., Cotoia, A., McLean, Kenneth A., Lizzi, V., Vovola, F., Bruzzaniti, P., Familiari, P., Lapolla, P., Marruzzo, G., Mingoli, A., Ribuffo, D., Cipriani, R., Contedini, F., Mckay, Siobhan C., Lauretta, M., Marchetti, C., Melotti, M., Pignatti, M., Pinto, V., Pizzigallo, A., Ricotta, F., Tarsitano, A., Catarzi, L., Abdulwahed, E. A., Glasbey, James, Mills, Emily C., Alshareea, E. A., Biala, M. I., Ghmagh, R. J., Ibrahim, A. F., Liew, Y. T., Alvarez, M. R., Arrangoiz, R., Cordera, F., Gómez-Pedraza, A., Soulé-Martínez, C. E., Becerril, O. S., Becerra, Gfc, Arkha, Y., Bechri, H., El Ouahabi, A., Oudrhiri, M., Benkabbou, A., Majbar, M., Mohsine, R., Souadka, A., Pellino, Gianluca, Lageju, N., Schreuder, W. H., Hardillo, J., de Bree, R., Schweitzer, D., Adeyeye, A. A., Enoch, E. E., Sholadoye, T. S. T. T., Wuraola, F., Oyelakin, O., Picciochi, Maria, Khokhar, M. I., Ayub, B., Walędziak, M., Szewczyk, M., Faria, C., Cardoso, P., Castro Silva, J., AlKharashi, E., Jelovac, D., Petrovic, M., Taylor, Elliott H., Sumrak, S., Asceric, R. R., Bojicic, J. M., Kovacevic, B. M., Krdzic, I. D., Milentijevic, M. A., Milutinovic, V. Z., Stefanovic, Z. B., Villacampa, J. M., Jiménez Carneros, V., Tiwari, Abhinav, Salazar Carrasco, A., Carabias Hernandez, A., Alonso Lamberti, L., León Ledesma, R., Jiménez Miramón, F. J., Jover Navalón, J. M., Garcia Quijada, J., Ramos Rodriguez, J., Valle Rubio, A., Vilaseca, I., Escartin, J., Estaire-Gomez, M., Padilla Valverde, D., Tousidonis, M., Lopez, F., Deandrés-Olabarria, U., Durán-Ballesteros, M., Fernández-Pablos, F., Ibáñez-Aguirre, F., Sanz-Larrainzar, A., Trout, Isobel M., Ugarte-Sierra, B., Di Martino, M., Prada, J., Jariod-Ferrer, U. M., Landaluce Olavarria, A., Rey-Biel, J., Díaz de Cerio, P., Sánchez Barrueco, A., Lindqvist, E. K., Sund, M., Venn, Mary L., Piantanida, R., Giger, R., Hool, S., Müller, S. A., Stoeckli, S. J., Simon, C., Toutounji, T., Al Assaf, A., Hammed, A. M., Hammed, S. M., Wilkin, Richard J. W., Mahfoud, M., Arikan, A., Yalkin, Ö, İflazoğlu, N., Isik, A., Leventoglu, S., Aydemir, L., Basaran, B., Sen, C., Comert-Ulusan, M., Ganly, Ian, Saracoglu, K. T., Saracoglu, A., Mantoglu, B., Kucuk, G., Aygun, N., Baran, E., Tanal, M., Eray Tufan, A., Uludag, M., Gürkan Yetkin, S., Yigit, B., Calik, B., Demirli Atici, S., Kaya, T., Sikakulya, F. K., Abdel-Galil, K. M. A. H., Lowe, T., Durrani, A. J., Habeeb, A., Irune, E., Luke, L., Masterson, L., Murphy, S. H., Segaren, N., Walker, C., Waseem, S., Jones, T. M., Loh, C., Pringle, S., Schache, A. G., Shaw, R. J., Stenhouse, J., Armstrong, M., Sood, S., Sutton, D., Thomas, S., Clarke, P., Winter, S. C., Hislop, S., Counter, P. R., Ghazali, N., Lloyd, C., Prabhu, V., Godden, D., Whitley, S., Butler, C., Nash, R., El-Boghdadly, K., Fry, A., Ho, Michael W. S., Niziol, R., De, M., Gill, C. K., Crank, S., Mace, A. D., Ho, M., Mair, M., Kothari, P., Homer, J., Sainuddin, S., Nankivell, Paul, Egan, R. J., Kittur, M., Burgess, C., O'Hara, J., Manickavasagam, J., McDonald, C., Burrows, S., Java, K. R., Katre, C., Ahmed, A., Biel, Juan Rey, Siddique, H., King, E., Ramchandani, P., Naredla, P. R., Brennan, P., Ringrose, T., Schmidt, F., Mak, J. K. C., Nankivell, P., Parmar, S., Batstone, Martin, Sharma, N., Douglas, C., McCaul, J., Dhanda, J., Kyzas, P., Vassiliou, L., Kumar, A., Husband, A., Hulbert, J., Ingrams, D., Parkin, R., Varley, I., Gahir, D., George, A., Zakai, D., Bater, M., Surwald, C., Devlin, B., Simon, Christian, Leonard, C. G., Pigadas, N., Snee, D., Singh, R. P., Hyde, N. C., Paley, M., Cocks, H., Wilson, A., Choi, D., Kerawala, C. J., Riva, F., Dickason, A., Semple, C. J., Schilling, C., Walton, G., Rees-Stoner, O., Scott, N., Nixon, I. J., Tighe, D., Abbott, Tom E. F., Mattine, S., Chu, M. M. H., Pothula, V., Lee, W., Brown, L., Ganly, I., Alpert, N., Illezeau, C. N., Miles, B., Rapp, J., Adamina, Michel, Taioli, E., Azam, M. T., Choudhry, A. J., Marx, W., Stein, J., Ying, Y., Gross, N. D., Almasri, M., Joshi, R., Kulkarni, G., Ademuyiwa, Adesoji O., Marwan, H., Mehdi, M., Sumer, B., Consorti, Giuseppe, Agarwal, Arnav, Alameer, Ehab, Alderson, Derek, Alakaloko, Felix, Albertsmeiers, Markus, Alser, Osaid, Alshaar, Muhammad, Alshryda, Sattar, Arnaud, Alexis P., Augestad, Knut Magne, Ayasra, Faris, Azevedo, José, Bankhead-Kendall, Brittany K., Barlow, Emma, Nankivell, Paul C., Blanco-Colino, Ruth, Brar, Amanpreet, Breen, Kerry A., Bretherton, Chris, Buarque, Igor Lima, Burke, Joshua, Caruana, Edward J., Chaar, Mohammad, Chakrabortee, Sohini, Christensen, Peter, Cox, Daniel, Cukier, Moises, Cunha, Miguel F., Davidson, Giana H., Desai, Anant, Di Saverio, Salomone, Drake, Thomas M., Edwards, John G., Elhadi, Muhammed, Collaborative, COVIDSurg, COVIDSurg Collaborative, EuroSurg, European Soc Coloproctology ESCP, Global Initiative Childrens Surg G, GlobalSurg, GlobalPaedSurg, ItSURG, PTSurg, SpainSurg, Italian Soc Colorectal Surg SICCR, Assoc Surg Training ASiT, Irish Surg Res Collaborative ISRC, Transatlantic Australasian Retrope, Italian Soc Surg Oncology SICO, Schache, Andrew G, Winter, Stuart C, Glasbey, Jame, Nankivell, Paul C, Simoes, Joana F F, Benson, Ruth A, Dawson, Brett E, Evans, Jonathan P, Glasbey, James C, Gujjuri, Rohan R, Jones, Conor S, Kamarajah, Sivesh K, Khaw, Rachel A, Keatley, James M, Mann, Harvinder S, Marson, Ella J, McLean, Kenneth A, Mckay, Siobhan C, Mills, Emily C, Taylor, Elliott H, Trout, Isobel M, Venn, Mary L, Wilkin, Richard J W, Ho, Michael W S, Abbott, Tom E F, Ademuyiwa, Adesoji O, Albertsmeiers, Marku, Arnaud, Alexis P, Ayasra, Fari, Bankhead-Kendall, Brittany K, Breen, Kerry A, Bretherton, Chri, Caruana, Edward J, Cukier, Moise, Cunha, Miguel F, Davidson, Giana H, Saverio, Salomone Di, Drake, Thomas M, Edwards, John G, Edward Fitzgerald, J, Griffiths, Ewen A, Harrison, Ewen M, Hutchinson, Peter J, Jenkinson, Michael D, Kaafarani, Haytham M A, Kolias, Angelo, Lederhuber, Han, Löffler, Markus W, Mashbari, Hassan N, Mazingi, Denni, Mohan, Helen M, Ng-Kamstra, Joshua S, Niquen, Milagro, Pinkney, Thomas D, Roslani, April C, Schnitzbauer, Andreas A, Seyi-Olajide, Justina O, Stewart, Grant D, Tsoulfas, Georgio, van Ramshorst, Gabrielle H, Warren, Oliver J, Boccalatte, L A, Batstone, M, Hodge, R, Abeloos, J, De Backer, T, De Ceulaer, J, Dick, C, Diez-Fraile, A, Lamoral, P, Spaas, C, Schrijvers, D L A L, Willemse, E B M, Faris, C, Maariën, S, Van Haesendonck, G, Van Laer, C, Deron, P, Abdallah, E A, Carvalho, G B, Kowalski, L, Vartanian, J, Gatti, A P, Nardi, C N, Oliva, R N L, Salem, M C, Cheng, D, MacNeil, D, Martin, J, Mayer, R, Groot, G, Acosta, L, Mejia, M, Perez, C J, Lorencin, M, Luksic, I, Mamic, M, Ashoush, F M, Osman, N A, Safwat Shahine, M, Eldaly, A, Elfiky, M M A, Amin, A, Elmorsi, R, Refky, B, Essa, M M, Mengistu, G Mengesha, Dakpé, S, Boucher, S, Ballouhey, Q, Laloze, J, Usseglio, J, Hoffmann, C, Gregoire, V, Lallemant, B, Blaurock, M, Reim, D, Boehm, A, Guntinas-Lichius, O, Hölzle, F, Modabber, A, Winnand, P, Kleeff, J, Lorenz, K, Ronellenfitsch, U, Schneider, R, Betz, C S, Böttcher, A, Busch, C, Möckelmann, N, Inhestern, J M, Greve, J, Hoffmann, T K, Laban, S, Vahl, J M, Agyeman-Prempeh, A, Aning, D, Barnor, I, Darko-Asante, R, Dzogbefia, M, Gaveh, V, Gyimah, D, Issahalq, M D, Konney, A, Poku, M, Adjeso, T, Akornor, E T, Amankwaa, W O, Antwi, D A, Apppiah-Thompson, P, Damah, M, Kumi, E O, Manan, L, Murphy, J P, Osei, L, Setuagbe, J, Arkadopoulos, N, Danias, N, Economopoulou, P, Frountzas, M, Kokoropoulos, P, Larentzakis, A, Michalopoulos, N V, Nastos, K, Parasyris, S, Pikoulis, E, Selmani, J, Sidiropoulos, T A, Vassiliu, P, Kalfountzos, C E, Chatziioannou, I, Corais, C, Gkrinia, E, Ntziovara, A, Saratziotis, A, Antoniadis, K, Orestis, O, Tatsis, D, Baili, E, Charalabopoulos, A, Liakakos, T, Schizas, D, Spartalis, E, Syllaios, A, Zografos, C, Aguilera-Arévalo, M, Misra, S, Pareek, P, Vishnoi, J, Chappity, P, Kar, M, Muduly, D K, Sultania, M, Agarwal, S, Garg, P K, Maharaj, D D, Majumdar, K S, Mishra, N, Poonia, D, Seenivasagam, R K, Singh, M P, Tiwari, A R, Penumadu, P, Rajan, S, Kumar, S, Raychowdhury, R, Ghodke, R, Barry, C, Callanan, D, Dias, A, Haung, L, Ionescu, A, Sheahan, P, Lennon, P, Fitzgerald, C, Mizrachi, A, Deganello, A, Pellini, R, Pichi, B, Lemma, F, Marino, M V, Bergonzani, M, Varazzani, A, Bussu, F, Perra, T, Piras, A, Porcu, A, Rizzo, D, Campisi, G, Cordova, A, Franza, M, Rinaldi, G, Toia, F, Gianni, A, Giannini, L, Gordini, L, Baldini, E, Conti, L, De Virgilio, A, Ferreli, F, Gaino, F, Mercante, G, Spriano, G, Ansarin, M, Chu, F, De Berardinis, R, Ietrobon, G, Tagliabue, M, Ionna, F, Baietti, A, Maremonti, P, Neri, F, Prucher, G, Ricci, S, Casaril, A, Nama, M, Cotoia, A, Lizzi, V, Vovola, F, Bruzzaniti, P, Familiari, P, Lapolla, P, Marruzzo, G, Mingoli, A, Ribuffo, D, Cipriani, R, Contedini, F, Lauretta, M, Marchetti, C, Melotti, M, Pignatti, M, Pinto, V, Pizzigallo, A, Ricotta, F, Tarsitano, A, Catarzi, L, Consorti, G, Abdulwahed, E A, Alshareea, E A, Biala, M I, Ghmagh, R J, Ibrahim, A F, Liew, Y T, Alvarez, M R, Arrangoiz, R, Cordera, F, Gómez-Pedraza, A, Soulé-Martínez, C E, Becerril, O S, Becerra, GFC, Arkha, Y, Bechri, H, El Ouahabi, A, Oudrhiri, M, Benkabbou, A, Majbar, M, Mohsine, R, Souadka, A, Lageju, N, Schreuder, W H, Hardillo, J, de Bree, R, Schweitzer, D, Adeyeye, A A, Enoch, E E, Sholadoye, T S T T, Wuraola, F, Oyelakin, O, Khokhar, M I, Ayub, B, Walędziak, M, Szewczyk, M, Faria, C, Cardoso, P, Castro Silva, J, AlKharashi, E, Jelovac, D, Petrovic, M, Sumrak, S, Asceric, R R, Bojicic, J M, Kovacevic, B M, Krdzic, I D, Milentijevic, M A, Milutinovic, V Z, Stefanovic, Z B, Villacampa, J M, Jiménez Carneros, V, Salazar Carrasco, A, Carabias Hernandez, A, Alonso Lamberti, L, León Ledesma, R, Jiménez Miramón, F J, Jover Navalón, J M, Garcia Quijada, J, Ramos Rodriguez, J, Valle Rubio, A, Vilaseca, I, Escartin, J, Estaire-Gomez, M, Padilla Valverde, D, Tousidonis, M, Lopez, F, Deandrés-Olabarria, U, Durán-Ballesteros, M, Fernández-Pablos, F, Ibáñez-Aguirre, F, Sanz-Larrainzar, A, Ugarte-Sierra, B, Di Martino, M, Prada, J, Jariod-Ferrer, U M, Landaluce Olavarria, A, Rey-Biel, J, Díaz de Cerio, P, Sánchez Barrueco, A, Lindqvist, E K, Sund, M, Piantanida, R, Giger, R, Hool, S, Müller, S A, Stoeckli, S J, Simon, C, Toutounji, T, Al assaf, A, Hammed, A M, Hammed, S M, Mahfoud, M, Arikan, A, İflazoğlu, N, Isik, A, Leventoglu, S, Aydemir, L, Basaran, B, Sen, C, Comert -Ulusan, M, Saracoglu, K T, Saracoglu, A, Mantoglu, B, Kucuk, G, Aygun, N, Baran, E, Tanal, M, Eray Tufan, A, Uludag, M, Gürkan Yetkin, S, Yigit, B, Calik, B, Demirli Atici, S, Kaya, T, Sikakulya, F K, Abdel-Galil, K M A H, Lowe, T, Durrani, A J, Habeeb, A, Irune, E, Luke, L, Masterson, L, Murphy, S H, Segaren, N, Walker, C, Waseem, S, Jones, T M, Loh, C, Pringle, S, Schache, A G, Shaw, R J, Stenhouse, J, Armstrong, M, Sood, S, Sutton, D, Thomas, S, Clarke, P, Winter, S C, Hislop, S, Counter, P R, Ghazali, N, Lloyd, C, Prabhu, V, Godden, D, Whitley, S, Butler, C, Nash, R, El-Boghdadly, K, Fry, A, Niziol, R, De, M, Gill, C K, Crank, S, Mace, A D, Ho, M, Mair, M, Kothari, P, Homer, J, Sainuddin, S, Egan, R J, Kittur, M, Burgess, C, O'Hara, J, Manickavasagam, J, McDonald, C, Burrows, S, Java, K R, Katre, C, Ahmed, A, Siddique, H, King, E, Ramchandani, P, Naredla, P R, Brennan, P, Ringrose, T, Schmidt, F, Mak, J K C, Nankivell, P, Parmar, S, Sharma, N, Douglas, C, McCaul, J, Dhanda, J, Kyzas, P, Vassiliou, L, Kumar, A, Husband, A, Hulbert, J, Ingrams, D, Parkin, R, Varley, I, Gahir, D, George, A, Zakai, D, Bater, M, Surwald, C, Devlin, B, Leonard, C G, Pigadas, N, Snee, D, Singh, R P, Hyde, N C, Paley, M, Cocks, H, Wilson, A, Choi, D, Kerawala, C J, Riva, F, Dickason, A, Semple, C J, Schilling, C, Walton, G, Rees-Stoner, O, Scott, N, Nixon, I J, Tighe, D, Mattine, S, Chu, M M H, Pothula, V, Lee, W, Brown, L, Ganly, I, Alpert, N, Illezeau, C N, Miles, B, Rapp, J, Taioli, E, Azam, M T, Choudhry, A J, Marx, W, Stein, J, Ying, Y, Gross, N D, Almasri, M, Joshi, R, Kulkarni, G, Marwan, H, Mehdi, M, and Sumer, B

Subjects
AcademicSubjects/MED00910, SARS-CoV-2, pandemic, COVID-19, safe healthcare delivery, education, Bjs/2, General Medicine, 030230 surgery, 3126 Surgery, anesthesiology, intensive care, radiology, 3. Good health, 03 medical and health sciences, fluids and secretions, 0302 clinical medicine, Head and Neck Neoplasms, 030220 oncology & carcinogenesis, parasitic diseases, Research Letter, Humans, head and neck cancer, Human medicine, AcademicSubjects/MED00010, COVID
Abstract

BJS open 5(6), zrab112 (2021). doi:10.1093/bjsopen/zrab112, Published by Oxford University Press, Oxford

Published
2021

2. Handicap-Recover Evolution Leads to a Chemically Versatile, Nucleophile-Permissive Protease

Author

Thomas Shafee, Gatti-Lafranconi P, Minter R, Hollfelder F, Hollfelder, Florian [0000-0002-1367-6312], and Apollo - University of Cambridge Repository

Subjects
nucleophilic catalysis, proteases, directed evolution, PA clan, tobacco etch virus, Communications, Uncategorized
Abstract

© 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Mutation of the tobacco etch virus (TEV) protease nucleophile from cysteine to serine causes an approximately 104-fold loss in activity. Ten rounds of directed evolution of the mutant, TEVSer, overcame the detrimental effects of nucleophile exchange to recover near-wild-type activity in the mutant TEVSerX. Rather than respecialising TEV to the new nucleophile, all the enzymes along the evolutionary trajectory also retained the ability to use the original cysteine nucleophile. Therefore the adaptive evolution of TEVSer is paralleled by a neutral trajectory for TEVCys, in which mutations that increase serine nucleophile reactivity hardly affect the reactivity of cysteine. This apparent nucleophile permissiveness explains how nucleophile switches can occur in the phylogeny of the chymotrypsin-like protease PA superfamily. Despite the changed key component of their chemical mechanisms, the evolved variants TEVSerX and TEVCysX have similar activities; this could potentially facilitate escape from adaptive conflict to enable active-site evolution.

Published
2021
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3. A Shorter Route to Antibody Binders $\textit{via}$ Quantitative $\textit{in vitro}$ Bead-Display Screening and Consensus Analysis

Author

Mankowska, SA, Gatti-Lafranconi, P, Chodorge, M, Sridharan, S, Minter, RR, Hollfelder, F, Hollfelder, Florian [0000-0002-1367-6312], and Apollo - University of Cambridge Repository

Subjects
molecular engineering, high-throughput screening, antibody therapy, biotechnology
Abstract

Affinity panning of large libraries is a powerful tool to identify protein binders. However, panning rounds are followed by the tedious re-screening of the clones obtained to evaluate binders precisely. In a first application of Bead Surface Display (BeSD) we show successful $\textit{in vitro}$ affinity selections based on flow cytometric analysis that allows fine quantitative discrimination between binders. Subsequent consensus analysis of the resulting sequences enables identification of clones that bind tighter than those arising directly from the experimental selection output. This is demonstrated by evolution of an anti-Fas receptor single-chain variable fragment (scFv) that was improved 98-fold $\textit{vs}$ the parental clone. Four rounds of quantitative screening by fluorescence-activated cell sorting of an error-prone library based on fine discrimination between binders in BeSD were followed by analysis of 200 full-length output sequences that suggested a new consensus design with a $K_d$ ~ 140pM. This approach shortens the time and effort to obtain high affinity reagents and its cell-free nature transcends limitations inherent in previous $\textit{in vivo}$ display systems.

Published
2016

5. Limnological and ecological methods: approaches, and sampling strategies for middle Xingu River in the area of influence of future Belo Monte Power Plant

Author

Tundisi, JG., Matsumura-Tundisi, T., Tundisi, JEM., Faria, CRL., Abe, D.S., Blanco, F., Rodrigues Filho, J., Campanelli, L., Sidagis Galli, C., Teixeira-Silva, V., Degani, R., Soares, FS., and Gatti Junior, P.

Subjects
ecosystem, limnologia, Xingu River, rios, limnology, Rio Xingu, ecossistemas, estratégias de amostragem, Belo Monte Power Plant, rivers, sampling strategies, UHE Belo Monte
Abstract

In this paper the authors describe the limnological approaches, the sampling methodology, and strategy adopted in the study of the Xingu River in the area of influence of future Belo Monte Power Plant. The river ecosystems are characterized by unidirectional current, highly variable in time depending on the climatic situation the drainage pattern an hydrological cycle. Continuous vertical mixing with currents and turbulence, are characteristic of these ecosystems. All these basic mechanisms were taken into consideration in the sampling strategy and field work carried out in the Xingu River Basin, upstream and downstream the future Belo Monte Power Plant Units. Resumo Neste trabalho os autores descrevem as abordagens limnológicas e a metodologia e estratégia de amostragem adotadas no estudo do Rio Xingu, na área de influência da futura Usina da Belo Monte (UHE Belo Monte). O ecossistema de rios é caracterizado por velocidade unidirecional de correntes, muitas flutuações de fluxos dependendo do clima do padrão de drenagem e do ciclo hidrológico. Mistura vertical permanente e turbulências ocorrem constantemente devido a correntes predominantes. Todos estes mecanismos gerais e específicos característicos do Rio Xingu, foram levados em consideração no plano de estudos limnológicos e amostragens desenvolvidas na área de influência de UHE Belo Monte a montante e a jusante do futuro empreendimento.

Published
2015

6. Spatial patterns of water quality in Xingu River Basin (Amazonia) prior to the Belo Monte dam impoundment

Author

Rodrigues-Filho, JL., Abe, DS., Gatti-Junior, P., Medeiros, GR., Degani, RM., Blanco, FP., Faria, CRL., Campanelli, L., Soares, FS., Sidagis-Galli, CV., Teixeira-Silva, V., Tundisi, JEM., Matsmura-Tundisi, T., and Tundisi, JG.

Subjects
Amazonian rivers, limnologia, hidrelétrica, limnology, rios amazônicos, bacia hidrográfica, hydropower dam, qualidade de água, water quality, watershed
Abstract

The Xingu River, one of the most important of the Amazon Basin, is characterized by clear and transparent waters that drain a 509.685 km2 watershed with distinct hydrological and ecological conditions and anthropogenic pressures along its course. As in other basins of the Amazon system, studies in the Xingu are scarce. Furthermore, the eminent construction of the Belo Monte for hydropower production, which will alter the environmental conditions in the basin in its lower middle portion, denotes high importance of studies that generate relevant information that may subsidize a more balanced and equitable development in the Amazon region. Thus, the aim of this study was to analyze the water quality in the Xingu River and its tributaries focusing on spatial patterns by the use of multivariate statistical techniques, identifying which water quality parameters were more important for the environmental changes in the watershed. Data sampling were carried out during two complete hydrological cycles in twenty-five sampling stations. The data of twenty seven variables were analyzed by Spearman's correlation coefficients, cluster analysis (CA), and principal component analysis (PCA). The results showed a high auto-correlation between variables (> 0.7). These variables were removed from multivariate analyzes because they provided redundant information about the environment. The CA resulted in the formation of six clusters, which were clearly observed in the PCA and were characterized by different water quality. The statistical results allowed to identify a high spatial variation in the water quality, which were related to specific features of the environment, different uses, influences of anthropogenic activities and geochemical characteristics of the drained basins. It was also demonstrated that most of the sampling stations in the Xingu River basin showed good water quality, due to the absence of local impacts and high power of depuration of the river itself. Resumo O rio Xingu, um dos mais importantes rios que constituem a bacia amazônica, é caracterizado por águas claras e transparentes que drenam uma bacia hidrográfica de 509,685 km2 com distintas condições hidrológicas, ecológicas e pressões antrópicas ao longo de seu percurso. Assim como nas demais bacias hidrográficas da Amazônia, os estudos no Xingu são raros. A eminente construção da Usina hidrelétrica de Belo Monte, a qual alterará as condições ambientais na bacia, demanda também a realização de estudos que gerem informações relevantes e que forneçam subsídios para um desenvolvimento mais equilibrado e igualitário região amazônica. Desta maneira, o objetivo deste trabalho foi analisar a qualidade de água no rio Xingu e em seus tributários focando em padrões espaciais na área de estudo pelo uso de técnicas de estatística multivariada, identificando quais parâmetros são mais relevantes nas condições de qualidade de água na bacia hidrográfica. As coletas de dados ocorreram durante dois ciclos hidrológicos completos em vinte e cinco estações de amostragem. Os dados de vinte e sete variáveis foram analisados pelos coeficientes de correlação de Spearman, análise Cluster (CA), e análise de componentes principais (PCA). Os resultados apontaram para um elevado contingente de variáveis com elevada correlação entre si (>0,7), sendo que estas altas variáveis foram retiradas das análises multivariadas por fornecerem informações ecológicas redundantes sobre o ambiente estudado. A CA resultou na formação de 6 agrupamentos, os quais foram observados por possuírem qualidade de água consideravelmente distintas pela PCA. Os resultados estatísticos permitiram identificar uma alta variação espacial da qualidade de água superficial na bacia estudada, sendo estas ocasionadas por especificidades do entorno, diferentes usos da água locais, influências de atividades antrópicas e características geoquímicas das bacias drenadas. Pode-se também evidenciar que em grande parte das estações de coleta na bacia do rio Xingu a qualidade de água superficial foi elevada, decorrente da ausência de impactos locais e do alto poder de depuração do rio.

Published
2015

8. Promiscuity, stability and cold adaptation of a newly isolated acylaminoacyl peptidase

Author

Pietro Gatti-Lafranconi, Electra Brunialti, Marina Lotti, Brunialti, E, GATTI LANFRANCONI, P, and Lotti, M

Subjects
Sporosarcina, Stereochemistry, Molecular Sequence Data, lac operon, Cold activity,Catalytic promiscuity,Flexibility, Acylaminoacyl peptidase, Enzyme isolation, Cleavage (embryo), Biochemistry, Substrate Specificity, Catalysis, Bacterial Proteins, Enzyme Stability, Thermal stability, Amino Acid Sequence, Enzyme kinetics, Cloning, Molecular, Psychrophile, chemistry.chemical_classification, Base Sequence, biology, Chemistry, General Medicine, biology.organism_classification, BIO/10 - BIOCHIMICA, Cold Temperature, Kinetics, Enzyme, Bacteria, Peptide Hydrolases
Abstract

We report on the characterisation of a member of the acylaminoacyl peptidase family, the first isolated from bacteria. The enzyme was obtained from the psychrophilic bacterium Sporosarcina psychrophila and shows the typical features of cold adaptation (low T m, optimal temperature of 40 °C, poor thermal stability). It was also tested for substrate specificity, effect of metals, temperature dependence and structure stability and revealed promiscuous catalytic activity on at least two chemically distinct substrates, with k cat/K m values for ester hydrolysis and acylamino acids cleavage of 1.7 × 10 4 s -1 M -1 and 6.2 × 10 3 s -1 M -1, respectively. Despite some properties cannot be explained with current models, results report on the relevance of structural and catalytic properties for the successful adaptation to cold temperatures. © 2011 Elsevier Masson SAS. All rights reserved.

Published
2011

9. Evolution of Stability in a Cold-Active Enzyme Elicits Specificity Relaxation and Highlights Substrate-Related Effects on Temperature Adaptation

Author

Antonino Natalello, Jürgen Pleiss, Pietro Gatti-Lafranconi, Marina Lotti, Sascha Rehm, Silvia Maria Doglia, GATTI LANFRANCONI, P, Natalello, A, Rehm, S, Doglia, S, Pleiss, J, and Lotti, M

Subjects
Protein Folding, Circular dichroism, Stereochemistry, Molecular Dynamics Simulation, Fluorescence, Protein Structure, Secondary, Substrate Specificity, Lactones, Molecular dynamics, temperature adaptation, Structural Biology, Enzyme Stability, differential scanning fluorimetry, Transition Temperature, Denaturation (biochemistry), directed evolution, Lipase, Pliability, Molecular Biology, Orlistat, biology, Chemistry, molecular dynamic, Circular Dichroism, Active site, Atmospheric temperature range, Directed evolution, BIO/10 - BIOCHIMICA, Adaptation, Physiological, Cold Temperature, Kinetics, Pseudomonas fragi, Structural Homology, Protein, biology.protein, Biophysics, Thermodynamics, Mutant Proteins, Protein folding, Directed Molecular Evolution
Abstract

Molecular aspects of thermal adaptation of proteins were studied by following the co-evolution of temperature dependence, conformational stability, and substrate specificity in a cold-active lipase modified via directed evolution. We found that the evolution of kinetic stability was accompanied by a relaxation in substrate specificity. Moreover, temperature dependence and selectivity turned out to be mutually dependent. While the wild-type protein was strictly specific for short-chain triglycerides (C4) in the temperature range 10-50 degrees C and displayed highest activity in the cold, its stabilized variant was able to accept C8 and C12 molecules and its selectivity was temperature dependent. We could not detect any improvement in the overall structural robustness of the mutant when the structure was challenged by temperature or chemical denaturants. There is, however, strong evidence for local stabilization effects in the active-site region provided by two independent approaches. Differential scanning fluorimetry revealed that the exposure of hydrophobic patches (as the active site is) precedes denaturation, and molecular dynamics simulations confirmed that stability was obtained by restriction of the mobility of the lid, a flexible structure that regulates the access to the enzyme active site and influences its stability. This reduction of lid movements is suggested to be accompanied by a concomitant increase in the mobility of other protein regions, thus accounting for the observed broadening of substrate specificity.

Published
2010

10. Concepts and tools to exploit the potential of bacterial inclusion bodies in protein science and biotechnology

Author

GATTI LANFRANCONI, PIETRO FRANCESCO, NATALELLO, ANTONINO, AMI, DILETTA, DOGLIA, SILVIA MARIA, LOTTI, MARINA, GATTI LANFRANCONI, P, Natalello, A, Ami, D, Doglia, S, and Lotti, M

Subjects
Inclusion Bodies, Protein Folding, Protein Conformation, electron and optical microscopie, FIS/07 - FISICA APPLICATA (A BENI CULTURALI, AMBIENTALI, BIOLOGIA E MEDICINA), stress response, native-like conformation, BIO/10 - BIOCHIMICA, Biochemistry, IB structural propertie, Recombinant Proteins, fourier transform infrared spectroscopy, Industrial Microbiology, Technology Transfer, Solubility, Stress, Physiological, biocatalysi, inclusion bodie, Escherichia coli, Animals, Humans, recombinant protein
Abstract

Cells have evolved complex and overlapping mechanisms to protect their proteins from aggregation. However, several reasons can cause the failure of such defences, among them mutations, stress conditions and high rates of protein synthesis, all common consequences of heterologous protein production. As a result, in the bacterial cytoplasm several recombinant proteins aggregate as insoluble inclusion bodies. The recent discovery that aggregated proteins can retain native-like conformation and biological activity has opened the way for a dramatic change in the means by which intracellular aggregation is approached and exploited. This paper summarizes recent studies towards the direct use of inclusion bodies in biotechnology and for the detection of bottlenecks in the folding pathways of specific proteins. We also review the major biophysical methods available for revealing fine structural details of aggregated proteins and which information can be obtained through these techniques.

Published
2011

11. How disorder influences order and viceversa: mutual effects in fusion proteins containing an intrinsically disordered and a globular protein

Author

Ilaria, Sambi, Pietro, Gatti-Lafranconi, Sonia, Longhi, Marina, Lotti, Sambi, I, Gatti Lafranconi, P, Longhi, S, and Lotti, M

Subjects
Kinetics, Protein Folding, Spectrometry, Fluorescence, disordered proteins, fusion proteins, GFP, measles virus, Protein Conformation, Circular Dichroism, Recombinant Fusion Proteins, Green Fluorescent Proteins, Proteins, Thermodynamics, Electrophoresis, Polyacrylamide Gel, BIO/10 - BIOCHIMICA, Algorithms
Abstract

Intrinsically disordered proteins (IDPs) are functional proteins either fully or partly lacking stable secondary and tertiary structure under physiological conditions that are involved in important biological functions, such as regulation and signalling in eukaryotes, prokaryotes and viruses. The function of many IDPs relies upon interactions with partner proteins, often accompanied by conformational changes and disorder-to-order transitions in the unstructured partner. To investigate how disordered and ordered regions interact when fused to one to another within the same protein, we covalently linked the green fluorescent protein to three different, well characterized IDPs and analyzed the conformational properties of the fusion proteins using various biochemical and biophysical approaches. We observed that the overall structure, compactness and stability of the chimeric proteins all differ from what could have been anticipated from the structural features of their isolated components and that they vary as a function of the fused IDP. © 2010 The Authors Journal compilation © 2010 FEBS.

Published
2010

12. Plasma-induced graft-polymerization of polyethylene glycol acrylate on polypropylene films: Chemical characterization and evaluation of the protein adsorption

Author

Pietro Gatti-Lafranconi, Elisa Grimoldi, Marina Lotti, Stefano Zanini, Silvia Maria Doglia, Anna Villa, Antonino Natalello, Claudia Colombo, Cristina Riccardi, Zanini, S, Riccardi, C, Grimoldi, E, Colombo, C, Villa, A, Natalello, A, GATTI LANFRANCONI, P, Lotti, M, and Doglia, S

Subjects
Materials science, Surface Properties, Analytical chemistry, Polyethylene glycol, Polypropylenes, Polyethylene Glycols, Biomaterials, chemistry.chemical_compound, Colloid and Surface Chemistry, Adsorption, Proteins adsorption, Fourier transform infrared spectroscopy, Polypropylene, chemistry.chemical_classification, Proteins, Polymer, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Graft copolymer, Acrylates, chemistry, Polymerization, Attenuated total reflection, Cold plasma, Biocompatibility, Nuclear chemistry, Protein adsorption
Abstract

This work deals with the optimization of argon plasma-induced graft-polymerization of polyethylene glycol acrylate (PEGA) on polypropylene (PP) films in order to obtain surfaces with a reduced protein adsorption for possible biomedical applications. To this end, we examined the protein adsorption on the treated and untreated surfaces. The graft-polymerization process consisted of four steps: (a) plasma pre-activation of the PP substrates; (b) immersion in a PEGA solution; (c) argon plasma-induced graft-polymerization; (d) washing and drying of the samples. The efficiency of these processes was evaluated in terms of the amount of grafted polymer, coverage uniformity and substrates wettability. The process was monitored by contact angle measurements, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), X-ray Photoelectron Spectroscopy (XPS) and atomic force microscopy (AFM) analyses. The stability of the obtained thin films was evaluated in water and in Phosphate Buffer Saline (PBS) at 37 °C. The adsorption of fibrinogen and green fluorescent protein (GFP) – taken as model proteins – on the differently prepared surfaces was evaluated through a fluorescence approach using laser scanning confocal microscopy with photon counting detection. After plasma treatments of short duration, the protein adsorption decreases by about 60–70% with respect to that of the untreated film, while long plasma exposure resulted in a higher protein adsorption, due to damaging of the grafted polymer.

Published
2010

13. Components of the E. coli envelope are affected by and can react to protein over-production in the cytoplasm

Author

Marina Lotti, Riccardo Villa, Pietro Gatti-Lafranconi, Villa, R, Lotti, M, Gatti Lafranconi, P, and Apollo - University of Cambridge Repository

Subjects
Vesicle-associated membrane protein 8, Membrane lipids, lcsh:QR1-502, Bioengineering, LIPOPOLYSACCHARIDE, Biology, Applied Microbiology and Biotechnology, lcsh:Microbiology, Inclusion bodies, Cell membrane, medicine, Protein biosynthesis, HEAT-SHOCK PROTEINS, OUTER-MEMBRANE PROTEINS, STRESS-RESPONSE, CELL-ENVELOPE, Secretory pathway, Research, AGGREGATION, BIO/10 - BIOCHIMICA, Cell biology, protein overexpression, stress response, bacterial envelope, medicine.anatomical_structure, Membrane protein, Biochemistry, ESCHERICHIA-COLI, Cytoplasm, LIPASE, OVEREXPRESSION, BACTERIAL INCLUSION-BODIES, Biotechnology
Abstract

Background Protein over-expression in bacteria is still the easiest, cheapest and therefore preferred way to obtain large amounts of proteins for industrial and laboratory scale preparations. Several studies emphasized the importance of understanding cellular and molecular mechanisms triggered by protein over-production in order to obtain higher yield and better quality of the recombinant product. Almost every step leading to a fully functional polypeptide has been investigated, from mRNA stability to the role of molecular chaperones, from aggregation to bottlenecks in the secretory pathway. In this context, we focused on the still poorly addressed relationship between protein production in the cytoplasm and the bacterial envelope, an active and reactive cell compartment that controls interactions with the environment and several major cellular processes. Results available to date show that the accumulation of foreign proteins in the cytoplasm induces changes in the membrane lipids and in the levels of mRNAs for some membrane proteins. However, a direct connection between membrane protein expression levels and soluble/aggregated protein accumulation in the cytoplasm has never been reported. Results By the use of a combined physiological and proteomic approach, we investigated the effects on the cell membrane of E. coli of the overexpression of two recombinant proteins, the B. cepacia lipase (BCL) and the green fluorescent protein (GFP). Both polypeptides are expressed in the cytoplasm at similar levels but GFP is fully soluble whereas inactive BCL accumulates in inclusion bodies. Growth and viability of the transformed cells were tested in the presence of different drugs. We found that chloramphenycol preferentially inhibited the strain over-producing GFP while SDS was more effective when BCL inclusion bodies accumulated in the cytoplasm. In contrast, both proteins induced a similar response in the membrane proteome, i.e. increased levels of LamB, OmpF, OmpA and TolC. Under all tested conditions, the lipopolysaccharide was not affected, suggesting that a specific rather than a generalized rearrangement of the envelope was induced. Conclusion Taking together physiological and biochemical evidence, our work indicates that the E. coli envelope can sense protein over-expression in the cytoplasm and react by modulating the abundance of some membrane proteins, with possible consequences on the membrane traffic of small solutes, i.e. nutrients, drugs and metabolites. Such a response seems to be independent on the nature of the protein being over-expressed. On the other hand both our data reported herein and previous results indicate that membrane lipids may act as a second stress sensor responsive to the aggregation state of the recombinant protein and further contribute to changes in cellular exchanges with the environment.

Published
2009

14. Effects of recombinant protein misfolding and aggregation on bacterial membranes

Author

Marina Lotti, Silvia Maria Doglia, Diletta Ami, Antonino Natalello, A. de Marco, Pietro Gatti-Lafranconi, Tina Schultz, Ami, D, Natalello, A, Schultz, T, Gatti Lafranconi, P, Lotti, M, Doglia, S, and De Marco, A

Subjects
Protein Folding, Vesicle-associated membrane protein 8, Cell Membrane Permeability, Recombinant protein, Membrane permeability, Recombinant Fusion Proteins, Membrane lipids, Green Fluorescent Proteins, Biophysics, Protein aggregation, Biology, Biochemistry, Analytical Chemistry, Green fluorescent protein, Cell membrane, Membrane Lipids, Genes, Reporter, Spectroscopy, Fourier Transform Infrared, Escherichia coli, medicine, Molecular Biology, Glutathione Transferase, Cell Membrane, Fourier transform infrared spectroscopy, Membrane lipid, beta-Galactosidase, BIO/10 - BIOCHIMICA, Cell biology, Stress metabolism, Oxidative Stress, medicine.anatomical_structure, Small chaperone, Protein folding, Intracellular
Abstract

The expression of recombinant proteins is known to induce a metabolic rearrangement in the host cell. We used aggregation-sensitive model systems to study the effects elicited in Escherichia coli cells by the aggregation of recombinant glutathione-S-transferase and its fusion with the green fluorescent protein that, according to the expression conditions, accumulate intracellularly as soluble protein, or soluble and insoluble aggregates. We show that the folding state of the recombinant protein and the complexity of the intracellular aggregates critically affect the cell response. Specifically, protein misfolding and aggregation induce changes in specific host proteins involved in lipid metabolism and oxidative stress, a reduction in the membrane permeability, as well as a rearrangement of its lipid composition. The temporal evolution of the host cell response and that of the aggregation process pointed out that the misfolded protein and soluble aggregates are responsible for the membrane modifications and the changes in the host protein levels. Interestingly, native recombinant protein and large insoluble aggregates do not seem to activate stress markers and membrane rearrangements.

Published
2009

15. Unscrambling thermal stability and temperature adaptation in evolved variants of a cold-active lipase

Author

Lilia Alberghina, Marina Lotti, Pietro Gatti-Lafranconi, Serena M. Caldarazzo, Alessandra Villa, Gatti Lafranconi, P, Caldarazzo, S, Villa, A, Alberghina, L, and Lotti, M

Subjects
Cold-activity, Hot Temperature, Protein Conformation, Molecular Sequence Data, Biophysics, Biochemistry, Bacterial Proteins, Structural Biology, Pseudomonas fragi, Enzyme Stability, Genetics, Thermal stability, Lid, Amino Acid Sequence, Lipase, Molecular Biology, Protein secondary structure, biology, Chemistry, Wild type, Cell Biology, Atmospheric temperature range, biology.organism_classification, Directed evolution, BIO/10 - BIOCHIMICA, Cold Temperature, Mutation, biology.protein, Specific activity, Directed Molecular Evolution, Stability
Abstract

Directed evolution by error-prone PCR was applied to stabilize the cold-active lipase from Pseudomonas fragi (PFL). PFL displays high activity at 10 °C, but it is highly unstable even at moderate temperatures. After two rounds of evolution, a variant was generated with a 5-fold increase in half-life at 42 °C and a shift of 10 °C in the temperature optimum, nevertheless retaining cold-activity. The evolved lipase displayed specific activity higher than the wild type enzyme in the temperature range 29¿42 °C. Biophysical measurements did not indicate any obvious difference between the improved variant and the wild type enzyme in terms of loss of secondary structure upon heat treatment, nor a shift in the apparent melting temperature. 2008 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Published
2008

16. Fourier transform infrared spectroscopy analysis of the conformational quality of recombinant proteins within inclusion bodies

Author

Silvia Maria Doglia, Marina Lotti, Pietro Gatti-Lafranconi, Antonino Natalello, Diletta Ami, Doglia, S, Ami, D, Natalello, A, Gatti Lafranconi, P, and Lotti, M

Subjects
Protein Folding, Protein Conformation, Kinetics, FIS/07 - FISICA APPLICATA (A BENI CULTURALI, AMBIENTALI, BIOLOGIA E MEDICINA), Applied Microbiology and Biotechnology, Inclusion bodies, law.invention, Protein structure, law, Spectroscopy, Fourier Transform Infrared, Fourier transform infrared spectroscopy, Solubility, Fourier transform infrared spectroscopy · Inclusion bodies · Recombinant proteins · Solubility, Inclusion Bodies, Chemistry, General Medicine, BIO/10 - BIOCHIMICA, Recombinant Proteins, FIS/01 - FISICA SPERIMENTALE, Biochemistry, Solubilization, Recombinant DNA, Molecular Medicine, Electrophoresis, Polyacrylamide Gel, Function (biology)
Abstract

The solubility of recombinant proteins produced in bacterial cells is considered a key issue in biotechnology as most overexpressed polypeptides undergo aggregation in inclusion bodies, from which they have to be recovered by solubilization and refolding procedures. Physiological and molecular strategies have been implemented to revert or at least to control aggregation but they often meet only partial success and have to be optimized case by case. Recent studies have shown that proteins embedded in inclusion bodies may retain residual structure and biological function and question the former axiom that solubility and activity are necessarily coupled. This allows for a switch in the goals from obtaining soluble products to controlling the conformational quality of aggregated proteins. Central to this approach is the availability of analytical methods to monitor protein structure within inclusion bodies. We describe here the use of Fourier transform infrared spectroscopy for the structural analysis of inclusion bodies both purified from cells and in vivo. Examples are reported concerning the study of kinetics of aggregation and structure of aggregates as a function of expression levels, temperature and co-expression of chaperones.

Published
2008

17. The lid is a structural and functional determinant of lipase activity and selectivity

Author

Francesco Secundo, Thorsten Eggert, Pietro Gatti-Lafranconi, Karl-Erich Jaeger, Stefania Brocca, Michael Puls, Chiara Tarabiono, Marina Lotti, Giacomo Carrea, Secundo, F, Carrea, G, Tarabiono, C, Gatti Lafranconi, P, Brocca, S, Lotti, M, Jaeger, K, Puls, M, and Eggert, T

Subjects
specificity, Bioengineering, Bacillus subtilis, Biochemistry, Catalysis, Candida rugosa lipase isoform 1, Pseudomonas fragi, enantioselectivity, Pseudotnonas fragi lipase, Lipase, domain swapping, Site-directed mutagenesis, Thermostability, chemistry.chemical_classification, biology, Chemistry, Process Chemistry and Technology, Active site, biology.organism_classification, BIO/10 - BIOCHIMICA, eye diseases, Candida rugosa, Enzyme, temperature stability, biology.protein, Bacillus subtilis lipase A, site-directed mutagenesi
Abstract

In several lipases access to the enzyme active site is regulated by the position of a mobile structure named the lid. The role of this region in modulating lipase function is reviewed in this paper analysing the results obtained with three different recombinant lipases modified in the lid sequence: Candida rugosa lipase isoform 1 (CRL1), Pseudomonas fragi lipase (PFL) and Bacillus subtilis lipase A (BSLA). A CRL chimera enzyme obtained by replacing its lid with that of another C. rugosa lipase isoform (CRL1LID3) was found to be affected in both activity and enantioselectivity in organic solvent. Variants of the PFL protein in which three polar lid residues were replaced with amino acids strictly conserved in homologous lipases displayed altered chain length preference profile and increased thermostability. On the other hand, insertion of lid structures from structurally homologous enzymes into BSLA, a lipase that naturally does not possess such a lid structure, caused a reduction in the enzyme activity and an altered substrate specificity. These results strongly support the concept that the lid plays an important role in modulating not only activity but also specifity, enantioselectivity and stability of lipase enzymes. (c) 2006 Elsevier B.V. All rights reserved.

Published
2006

18. Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy

Author

Antonino Natalello, Diletta Ami, Marina Lotti, Pietro Gatti-Lafranconi, Silvia Maria Doglia, Ami, D, Natalello, A, Gatti Lafranconi, P, Lotti, M, and Doglia, S

Subjects
Kinetics, Biophysics, inclusion body, Protein aggregation, medicine.disease_cause, Biochemistry, Inclusion bodies, Protein Structure, Secondary, law.invention, protein aggregation, Structural Biology, law, Spectroscopy, Fourier Transform Infrared, Genetics, medicine, Escherichia coli, lipase, FT-IR microspectroscopy, Lipase, Molecular Biology, Protein secondary structure, Inclusion Bodies, biology, Chemistry, Cell Biology, BIO/10 - BIOCHIMICA, Recombinant Proteins, Crystallography, kinetics of recombinant protein production, FIS/01 - FISICA SPERIMENTALE, Recombinant DNA, biology.protein, Inclusion (mineral)
Abstract

The aggregation of a recombinant lipase as inclusion bodies (IBs) was studied directly within intact Escherichia coli cells by FT-IR microspectroscopy. Through this approach, it was possible to monitor in real time the different kinetics of IB formation at 37 and 27 degrees C, in excellent agreement with the results of the SDS-PAGE analysis. Furthermore, insights on the residual native-like structure of the expressed protein within IB - both isolated and inside cells - were obtained by the secondary structure analysis of the Amide I band in the IB FT-IR spectra. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Published
2005

19. Directed evolution of a cold-active lipolytic enzyme

Author

Pietro Gatti-Lafranconi, Serena Caldarazzo, Marina Lotti, Lilia Alberghina, Gatti Lafranconi, P, Caldarazzo, S, Alberghina, L, and Lotti, M

Subjects
Biochemistry, Lipolytic enzyme, Chemistry, Bioengineering, General Medicine, Directed evolution, cold-active, BIO/10 - BIOCHIMICA, Applied Microbiology and Biotechnology, Biotechnology
Published
2007

20. Mutations in the 'lid' region affect chain length specificity and thermostability of a Pseudomonas fragi lipase

Author

Marina Lotti, Piercarlo Fantucci, Lilia Alberghina, Claudia Alquati, Gianluca Santarossa, Luca DeGioia, Pietro Gatti Lafranconi, Santarossa, G, Gatti Lafranconi, P, Alquati, C, DE GIOIA, L, Alberghina, L, Fantucci, P, and Lotti, M

Subjects
Models, Molecular, Protein Denaturation, Molecular Sequence Data, Biophysics, Biology, Biochemistry, Substrate Specificity, Serine, chemistry.chemical_compound, Structural Biology, Pseudomonas fragi, Enzyme Stability, Genetics, Amino Acid Sequence, Lipase, Molecular Biology, Triglycerides, Thermostability, CHIM/03 - CHIMICA GENERALE E INORGANICA, chemistry.chemical_classification, Binding Sites, Triglyceride, Hydrolysis, Wild type, Temperature, Cell Biology, biology.organism_classification, BIO/10 - BIOCHIMICA, Amino acid, Protein Structure, Tertiary, Kinetics, chemistry, Amino Acid Substitution, Glycine, Mutation, biology.protein, Chain length selectivity, Caprylates, Sequence Alignment
Abstract

The cold-adapted Pseudomonas fragi lipase (PFL) displays highest activity on short-chain triglyceride substrates and is rapidly inactivated at moderate temperature. Sequence and structure comparison with homologous lipases endowed with different substrate specificity and stability, pointed to three polar residues in the lid region, that were replaced with the amino acids conserved at equivalent positions in the reference lipases. Substitutions at residues T137 and T138 modified the lipase chain-length preference profile, increasing the relative activity towards C8 substrates. Moreover, mutations conferred to PFL higher temperature stability. On the other hand, replacement of the serine at position 141 by glycine destabilized the protein.

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