Novel insights into the action of antimicrobial agents against human pathogens

215 Pages·2013·5.43 MB·English

Novel insights into the action of antimicrobial
agents against human pathogens
Ana Filipa Nogueira Tavares
O
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I III IV
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CO H2O
CO-RM
O •-
2 Nitrofurans
activated Nitrofurans
SOD
Fe2+ + H2O2 OH• NtrA
GSNHOH GSNO
Fe-S
DNA
proteins
Protein Protein
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SH SH S S
Thiol-containing
proteins
Dissertation presented to obtain the Ph.D degree in Biochemistry
Instituto de Tecnologia Química e Biológica | Universidade Nova de Lisboa
Oeiras,
April, 2013

Novel insights into the action
of antimicrobial agents against
human pathogens
Ana Filipa Nogueira Tavares
Dissertation presented to obtain the Ph.D degree in Biochemistry
Instituto de Tecnologia Química e Biológica | Universidade Nova de Lisboa
Supervisor: Dr. Lígia M. Saraiva
Oeiras, April 2013
From left to the right: Maria de Fátima Lopes, Mónica Oleastro, Lígia Saraiva, Ivo
Boneca, Mário Ramirez, Ana Filipa Tavares and Carlos Romão
2nd April 2013

Second Edition, April 2013

Molecular Genetics of Microbial Resistance Laboratory
Instituto de Tecnologia Química e Biológica
Universidade Nova de Lisboa
Av. da República (EAN)
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Acknowledgements

I would like to formally express my sincere gratitude to the people that
contributed and supported my work:

To Dr. Lígia Saraiva for receiving me in her laboratory and accepting to be my
supervisor. Thank you for your constant support, encouragement and endless
capacity to always see the bright side, even in hard and more challenging
moments. For the trust and confidence in me, for your friendship and 24 h
availability whenever I needed to ask a question or communicate the result of
an experiment! Thank you for your great advices and for the careful revision of
this thesis. A big thank you for everything!!!

To Prof. Miguel Teixeira, for the collaboration on the work presented in
Chapter 4, especially for performing the EPR experiments, and for the very
interesting and helpful discussions.

To Prof. Carlos Romão, for the collaboration on the work presented in Chapter
4, for the enthusiastic discussions and all the lessons about CO-Releasing
Molecules chemistry.

To Dr. Mónica Oleastro, from Instituto Dr. Ricardo Jorge, for the collaboration
on the work presented in Chapter 5, for receiving me in her laboratory and for
the helpful advices. I would like also to thank João Benoliel for his help during
my stay at this institute.

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To Dr. Mariana Pinho and Dr. Ilda Sanches for being part of my scientific
thesis commission, for their suggestions and helpful advices.

To Dr. Colin McVey, from Structural Genomics lab at ITQB for reviewing the
abstract of this thesis, accordingly with ITQB intern rules.

To all past and present colleagues of MGMR lab´s for making it such an
enjoyable and pleasant place to work. A very special thank you to Lígia Nobre,
for all the help during the course of this work, for always have time to listen
my new ideas or results, for the valuable discussions and for being a good
friend. To Marta Justino, for the great discussions, all the help in statistical
analyses and for always willing to give me new ideas and suggestions. To
Margarida Parente, for the help provided in the last part of the experimental
work. To Susana Lobo, for her support, friendship and also for the help with
the cover of this thesis! To Mafalda Figueiredo, for her companionship and for
being a great friend. To Sara Sousa, for her friendship and good moments. To
Joana Baptista, for being a special friend, for all the funny moments inside and
outside the lab, for being my “shoulder friend” when I need to laugh or cry!
To former members of the group, especially to Cláudia Almeida, for her help
in my early days at the lab, and to Vera Gonçalves, for the good talks and
laughs.
Thank you all for so many moments of happiness and so many experiences
shared in the last years.

To everyone in the third floor of ITQB, past and present people, for
contributing to the helpful and friendly environment.

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To Alfama members for theirs pleasantness, good talks and for providing CO-
Releasing Molecules. In special to João Seixas, for the collaboration on the
work presented in Chapter 4.

To all my colleagues in other labs at the institute, especially to those from the
2008 class of the ITQB PhD course and from the InTeraQB. Thank you for
providing me good moments, for the experiences changed and friendly words.

To my colleagues from University that became very especial friends:
Margarida Augusto, Joana Medeiros and Simão Luz, for all the fun and
friendship. To Inês Lima, for her comprehension and support, a true and
special friendship full of precious and unforgotten moments throughout these
10 years!

Aos meus amigos de sempre e a todos os outros, muito obrigada por todo o
apoio e amizade.

À minha família pelo vosso interminável e incondicional apoio e carinho e a
quem devo aquilo que sou.
Aos meus pais, obrigada por acreditarem em mim e me darem força para ser
sempre melhor e para tentar ir sempre mais longe.
À minha irmã Xana pela amizade, pelo apoio e por estares sempre presente
em todos os momentos da minha vida!
E aos meus avós, por me acompanharem e ajudarem durante toda a vida.
OBRIGADA a todos por serem o meu suporte para tudo na vida.

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Ao Pedro, por me teres sempre apoiado, pela paciência e compreensão
inesgotável. Obrigada por estares sempre ao meu lado em todos os momentos
e por me fazeres feliz!

Fundação para Ciência e Tecnologia for the financial support crucial for the
accomplishment of this work and by awarding a PhD grant
(SFRH/BD/38457/2007).

This thesis is dedicated to my family

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Thesis Outline 

This thesis comprises the research work performed at the Molecular 
Genetics of Microbial Resistance Laboratory from the Instituto de Tecnologia 
Química e Biológica, Universidade Nova de Lisboa under the supervision of Dr. 
Lígia M. Saraiva. 
The thesis is divided in three main parts: Part I consists of a general 
introduction organized in two chapters, one focusing on bacteria and their 
resistance to antibiotics and to the innate immune system, and a second one 
concerning an overview on the general aspects of carbon monoxide and its 
relation  with  biological  systems  and  bacteria.  Part  II  comprises  the 
experimental  results  obtained  during  this  work,  which  is  divided  in  three 
chapters  based  on  two  original  publications  and  one  manuscript  in 
preparation. Part III presents a general discussion of all the work performed in 
this thesis. 

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