NATIONAL INSTITUTES OF SCIENCE AND
TECHNOLOGY
INCT of the Biomedicine of the Brazilian Semiarid
PARTIAL REPORT
2009/2010
INSTITUTO DE BIOMEDICINA DO SEMI-ÁRIDO BRASILEIRO
COORDINATOR
Name: ALDO ÂNGELO MOREIRA LIMA, CPF: 09055339334, R.G.Nº: 527969 Orgão Expedidor: SSP-CE
E-Mail: alima@ufc.br
Address: Centro de Biomedicina, Faculdade de Medicina, Universidade Federal do Ceará, R Cel Nunes
de Melo 1315 – Rodolfo Teófilo – CEP 60.430-270
Porangabussu, Cidade: FORTALEZA, UF: CE
Tel: (85) 3366-8445 and 3366-8437
CORE MANAGER COMITEE:
Dr. Gerly A. C. Brito, Dr. Vietla Rao, Dr. Armênio A. Santos, Dr. Helena S. A. Monteiro, Dr. Dulciene
Queiroz
CONSULTANTS:
RICHARD L. GUERRANT, DANIEL SIFRIM, MANASSÉS C. FONTELES
HEADQUARTERS:
Universidade Federal do Ceará
Address: Av. da Universidade, 2853
CNPJ: 07.272.636/0001-31
Benfica, Fortaleza, CE
http:// www.ufc.br
Tel.: 33667300
Head of the Federal University of Ceara
Name: JESUALDO PEREIRA FARIAS; CPF: 112745143-04; RG Nº: 7311d Orgão Expedidor: CREA-CE
Address: Av. da Universidade, 2853
Benfica, Fortaleza, CE
CEP: 60020-181 Telefone: 3366 7305 / 7306 / 7307
E-Mail: greitor@ufc.br
Position: Rector
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Report – First Year [2009 – 2010] - INCT_Biomedicina / UFC
The National Institute of Biomedicine of the Brazilian SemiArid (INCT_IBISAB),
officially established and funded in 16/1/2009, has as general goal to pursue
excellence in basic, preclinical (in vivo and in vitro), and clinical research, for
development of biological markers, bio-products, as well as causal determinants
(genetic and environmental) of endemic and highly prevalent diseases in the regions of
the Brazilian semi-arid and Northeast. Our specific aim is to study and to develop
markers and bio-products for use in preventing, diagnosis and treatment of diseases
affecting the digestive and nervous systems, considered as highly prevalent in the
Brazilian semi-arid. Diseases like infant diarrhea, malnutrition, cancer and gastric ulcers
and their impact on weight and children's cognitive development will be targeted for
research in this Institute.
To achieve these goals, we conducted to date four (4) meetings of the Core
Manager Committee, whose main discussed topics and agreed decisions are described
below.
1. Core Manager Committee: meetings and decisions
The inaugural meeting of the IBISAB’s Core Manager Committee was performed on
06/Mar/2009 in Fortaleza with the presence of the following members: Aldo A.M. Lima,
Armenian Santos, Lucia Libanez, Dulciene Queiroz, Otoni C. Valle, Reinaldo B. Oriá, Pedro
Magalhães, Gerly A.C. Brito, Francisco Gondim and Noelia L. Lima. The general coordinator,
Aldo Lima introduced the Core Manager Committee and assigned the coordinators for each
thematic, and summarized the main topics of the research plan. On the occasion, Prof. Pedro
Magalhaes received the position of supervisor/constructor of an internet home page for the
IBISAB, task that was shared with Mr. Francisco Sousa Junior, a member of the Safety Data
Management team of the IBISAB. Moreover, the annual meeting agenda for the Core Manager
Committee was decided, which was scheduled to the first Wednesday of the each month, in
every two month period. Prof. Armenio informed details about a recent meeting that occurred in
Brasilia with the presence of Prof. Luiz Antonio Barreto de Castro from the Secretary of
Programs and Policies for Research and Development, of the Ministry of Science and
Technology, along with representatives of the other National Institutes (INCTs), dedicated to
biomedical research in the country. In this meeting, the INCTs were invited to form the network
called SIBRATEC. The secretary requested a list of possible processes and/or drugs that could
be developed by the INCTs, since that would be of interest to companies and industries
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operating in Brazil. Participants were informed that available productivity of each researcher
should be incorporated in the IBISAB’s annual report. Research group demands should be
outlined by each thematic coordinator in order to build a list of priorities to be further established
by the steering committee. Prof. Dulciene suggested that those with greater access to
scholarship funding from other agencies should withdraw, as much as possible, to prevent
further use of the IBISAB budget, thus making possible the project implementation, as it was
designed.
In another meeting held on April 23, 2009, the following attendees were: Aldo A.M.
Lima, Armenio Santos, Lucia Libanez, Vietla Rao, Flávia Santos, Reinaldo B. Oriá, Pedro
Magalhães, Gerly A.C. Brito, Ronaldo Ribeiro and Marcellus Souza. The meeting consisted of
the following topics: i) Spreadsheet reports to predict budget expenditure on equipment and
funding available for each core thematic, complying with 20% tax cut according to CNPq
approved budget; ii) Laboratory Management for multi-users; iii) Personnel Training in the Multiuser laboratories; iv) Establishment of monthly research seminars at the School of Medicine
INCT and v) News of the Ministry of Science and Technology INCT reports.
On the next meeting held on January 20, 2010, the following attendees were: Aldo A.M.
Lima, Armenio Santos, Lucia Libanez and Pedro Magalhaes. The general coordinator, Aldo
Lima, reported an "e-mail” received from CNPq requiring annual INCT reporting. Research
productivity such as articles and patents should be listed by each thematic. Dr. Aldo Lima
presented predicted expenditures by INCT-IBISAB each thematic and also balance sheets and
expenditures of the first year. Dr. Aldo informed about the 14th Tropical Medicine Research
Center (TMRC) Meeting held during March, 4-6th, 2010 in Fortaleza and agreed with event
opening to all IBISAB thematic groups. Another discussed issue was the IBISAB available
scholarship funding, and Dr. Aldo Lima suggested the release of position announcement for
scholarships. Dr. Aldo Lima’s suggestion was accepted and each thematic was to receive a
post-doctoral fellowship as well as one technician stipend. Positions are to be selected
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according to curricula analysis and technical background. It was established that Prof. Armenio
Aguiar along with thematic coordinators should prepare an announcement draft to be forecast
nationally, and to perform the selection process.
On the last meeting held on April 26, 2010 were present: Aldo A.M. Lima, Reinaldo B.
Oriá, Pedro Magalhães, Armenio Santos, Alberto Melo and Lucia Libanez. The preliminary
version of the IBISAB first report was presented, soon to be sent to the CNPq. In this report
version was evident the relevant scientific production of the IBISAB-born groups.
Group
research development was evaluated in terms of scientific production, training of human
resources, knowledge and technology transfer (markers, bio-products and patents) and
education and scientific divulgation. Additionally, several changes in the IBISAB membership
research team were approved, such as the inclusion of the new members Prof. Antonio Aldo
Melo Filho (thematic IV) and Dra. Flávia Almeida Santos (thematic II). Thematic III coordination
change was approved having now Prof. Reinaldo B. Oriá, due to the former coordinator, Prof.
Carlos Maurício de Castro Costa, decease. Moreover, due to recent emigration for USA,
Professor Francisco A.A. Gondim and Dr. Gisele Ramos de Oliveira were excluded from the
Thematic III. Prof. Armenio Santos reported the outlined the IBISAB announcement for fellow
positions, which after being approved by the steering committee will be disclosed as early as in
May, 2010. Finally, the coordinator Aldo Lima launched the idea of hosting the next IBISAB
symposium, including guests from abroad, tentatively scheduled for March, 17-19th, 2011.
2. Cooperation activities between groups of INCT-IBISAB.
Due to severe shortage of technical and administrative staff in the Brazilian public
universities, the INCT-IBISAB coordination allocated the Clinical Research Unit (UPC)
employees to perform office and custom clearance of equipment and supplies requested by the
INCT-IBISAB researchers. Another academic cooperation was to share activities in the area of
informatics, as in the development of the Institute's internet home page, the creation of a
management and database system to support the researchers from each thematic, the creation
of a web system for management of multi-user laboratories and the acquisition and installation
of a video conference facility, all described below. It is noteworthy that the web system for
documentation is in advanced operation stage and the video conference room is already in
process of assembling, while the others are already available online.
Video conference room
In order to facilitate meetings, seminars and workshops among researchers from
different locations in Brazil and abroad, involved in the IBISAB studies, and to save costs with
lengthy travels, we planned to implement a video conferencing system that will allow
simultaneous internet connections up to four different locations worldwide. The complete
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system was already purchased and is now in the IBISAB facilities. It is scheduled for
functioning in the beginning of the second year INCT-IBISAB activities. This system will provide
the optimization of scientific activities involving either national or international research groups.
Website of IBISAB-INCT –– Home page
As requested by CNPq, a website was built under the address: <www.ibisab.ufc.br>.
The initial challenge was to create an interactive environment with unique visual identity without
being visually polluted and with the four different project themes presented. The subjects were
divided into sections and on the top menu the user could get access to the main laboratory
information as well as the scientific and technological publications of the Institute.
Moreover, database system implementation for document management is underway,
tool that will aid project researchers with their tasks, as can be seen in the figure below.
Website with access for a Multi-user Laboratory
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Increasingly sharing of scientific equipment in the academic environment is critical,
although difficulties in establishing the rational of equipment use may exist. To minimize these
setbacks, the INCT-IBISAB decided to build a digital agenda to access scientific sharing cores,
especially in those multi-user laboratories. The figure above depicts an example of on line
access to one of these environments. We believe that on line accessing through the network
will allow the dynamic registration and documentation and best performance of the shared
equipment users. The system is on line at: www.nempi.ufc.br, coordinated by Profs. Reinaldo
Oriá and Gerly A.C. Brito.
3. Cooperative activities between INCT-IBISAB with other institutions.
The dedication to academic studies, the relevance of scientific and aggregative work of
the INCT-IBISAB researcher members allowed, in this first year of activities, the establishment
of new scientific interactions between the IBISAB and other research networks, which are
described below.
DINTER – Inter-institucional PhD Course in Morphological Sciences between UFRJ and UFC
The researchers’ commitment and the INCT-IBISAB research infrastructure were
fundamental to the newly created inter-institutional PHD course in Morphological Sciences
involving the Federal University of Rio de Janeiro (UFRJ) and the Federal University of Ceará
(UFC), under the academic coordination of Prof. Vivaldo Moura Neto (UFRJ), Reinaldo Oriá
and Gerly Brito (UFC). The scientific activities will be held at the Department of Morphology of
UFC as well as in the IBISAB facilities.
Exploring the potential for nucleation of the Morphological Sciences Program of UFRJ,
the new course seeks to promote training skills in morph-functional sciences in order to awaken
new vocations to the academic life and trigger the formation new research groups interested in
the morphological studies of the digestive and nervous systems in the UFC School of Medicine
and in the INCT-IBISAB, thus improving the understanding and management of prevalent
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diseases among Semiarid populations. Furthermore, the partnership with the UFRJ will provide
the full use of infrastructure already available in the INCT-IBISAB enabling teaching careers in
Morphological Sciences in our Brazilian universities, especially those located in the University
campi spread in the Semiarid countryside. This is the first postgraduate program (PhD level) in
Morphological Sciences in the Brazilian North and Northeast regions that, certainly, will
contribute to minimizing regional disparities. For details, please see:
<http://www.prppg.ufc.br/index.php?option=com_content&view=article&id=689%3Acienciasmorfologicas&catid=141%3Adoutorado-interinstitucional&Itemid=56&lang=pt>
Research Network for Goat and Sheep Breeding for Prevention and Treatment of Infant
Diarrhea in the Brazilian SemiArid (RECODISA)
This network (RECODISA) is formed by researchers of three Brazilian institutions,
Federal University of Ceará (UFC), Ceará State University (UECE), University of Fortaleza
(UNIFOR), and an international institution, the University of California, Davis (UC Davis, USA)
with financial support from the Ministry of Science and Technology (FINEP Cross Order, Brazil),
having as main objective the identification of specific etiology and development of immune
compounds in the milk of transgenic goats for the prevention and treatment of childhood
diarrhea in the semiarid areas (for details see www.recodisa.ufc.br).
Motivated by the idea of offering immune compounds commonly found in human milk in
the milk of animals through the use of biotechnology and, then to benefit the health of children
at risk in the population of the semi-arid region of Brazil, RECODISA participate in the five postgraduate courses: (a) Northeast Biotechnology Network - RENORBIO (Level in Capes scale: 5
(ranging from 3 to 7); (b) Pharmacology - UFC (level 6); (c) Medical Sciences - UFC (level 5);
(d) Biotechnology (level 4; located in Sobral, a countryside city); and (e) the newly created interinstitutional PhD course in Morphological Sciences between UFC and UFRJ (level 6).
The impacts in terms of knowledge and technology transfer from INCT-IBISAB should
be significantly amplified by RECODISA. The specific objectives of RECODISA are: (a) to
determine the etiology of diarrhea in children 2-36 months of age seen at health facilities and to
compare their profile with healthy controls from the urban community in Fortaleza, CE, Brazil;
(2) To develop molecular methods for rapid and sensitive diagnosis of the etiologic agents and
to evaluate the genetic variation of virulence genes of the major causes of childhood diarrhea;
(3) to evaluate by a cross-sectional study the etiology of diarrhea in children aged 2-36 months
old in the semiarid region of Brazil; (4) To establish lines of heterozygous transgenic goats in
Brazil for the human lysozyme gene (HLZ) from UC Davis; (5) Purification and proteomic
analysis of HLZ expressed in the milk of transgenic goats, (6) Characterization of the in vitro
efficacy of the goat milk from HLZ transgenic lines against human pathogens commonly found
in Brazilian semiarid; (7) To conduct preclinical and clinical testing of purified HLZ from the milk
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of transgenic goats, (8) To generate heterozygous strains of transgenic goats with the gene of
human lactoferrin (HLF), through animal cloning procedures for somatic cell nuclear transfer
(SCNT); (9) Purification and proteomic analysis of HLF expressed in the milk of transgenic
goats; (10) characterization and in vitro efficacy of goat milk of the transgenic lines HLF against
human pathogens common to semiarid; (11) conduct pre-clinical tests and clinicians with HLF
and purified from the milk of transgenic goats for the HLF; (12) To generate heterozygous
strains of transgenic goats for peptide enriched with glutamine and arginine (PGA) by means of
animal cloning by SCNT; (13) Purification and proteomic analysis of PGA expressed in the milk
of transgenic goats; (14) characterization of in vitro efficacy of goat milk of the PGA transgenic
lines against human pathogens commonly found in Brazilian semiarid; (15) To conduct
preclinical and clinical testing studies with purified PGA and with milk of transgenic goats for the
PGA; (16) To create the breeding program for the establishment of homozygous lines of
transgenic goats for the genes of HLZ, HLF and PGA; (17) characterization of colonystimulating factor in human granulocytes (HG -CSF) expressed in the milk of transgenic goats;
(18) Cloning of Transgenic females for colony-stimulating factor in human granulocyte (hGCSF) by SCNT; (19) Purification and proteomic analysis of colony-stimulating factor in human
granulocytes ( hG-CSF) expressed in the milk of transgenic goats, and, finally, (20) To conduct
preclinical and clinical testing studies with purified hG-CSF.
In course of the first year we began the study on childhood diarrhea, covering eight
cities located in semiarid with over fifty thousand inhabitants. Moreover, it has been developed
two transgenic bio-products: goat milk with human lysozyme (HLZ) and goat milk with the
colony-stimulating factor in human granulocytes. Such products are now undergoing tests in
vitro and in vivo to demonstrate its feasibility and biological effectiveness for, then, to initiate the
preclinical and clinical testing (Castro LA & Barros AK. Incentives for Brazilian health biotech.
Nat Biotechnol . 27 (4) :317-8, 2009 and DR Brundige et al. Consumption of pasteurized human
lysozyme transgenic goats' milk alters serum metabolite profile in young pigs. Transgenic Res
2009 Oct 22.). Our outlook for the second year is quite encouraging given the initial success in
cloning these two byproducts, milk with HLZ (developed and produced at UC Davis) and milk
with hG-CSF (developed and produced in State University of Ceara, UECE).
International Network for the Study of Infant Diarrhea and Malnutrition (MAL_ED)
This network MAL_ED (Malnutrition_Enteric Diseases) was recently assembled upon
the support of the Bill and Melinda Gates Foundation for the global study of malnutrition and
infant diarrhea in the following countries: USA, Brazil, Bangladesh, South Africa, Tanzania,
India, Pakistan, Nepal and Peru. Among the main organizers of the proposed international
network, are the U.S., Bangladesh and Brazil, through this IBISAB (for details see the Internet
www.upcibimed.ufc.br).
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So far the INCT-IBISAB already developed and / or is developing several bio-markers
and molecular probes for specific diagnosis of causative agents of infantile diarrhea, biomarkers for assessing intestinal inflammation, such as lactoferrin and others, as well as
molecular probes, lactulose and mannitol, to determine the intestinal absorption area,
permeability and intestinal damage, and are now IBISAB international reference for some of
these products.
The project will establish a network of sites (multicenter) in developing countries in
partnership with the United States for better understanding the risk factors for malnutrition,
enteric diseases and their consequences for health, including the deficits in child development.
After all, one in five children in developing countries is malnourished and marginal nutrition is
associated with more than half of all deaths in children around the world. This project will carry
out epidemiological studies (longitudinal and case-control) at multiple sites to assess the
correlation level among: (a) infection by enteric pathogens, (2) intestinal barrier function
changes, (3) immune response to vaccines, and (4) Malnutrition with deficits in child
development.
Although some of these associations are already known, there are still few quantitative
and qualitative data on the specific etiologic agents, incidence by age, co-infection status and
further functional studies of the gastrointestinal tract with specific nutritional assessment - key
data to establish the target for optimal interventions. Moreover, we will formulate models of
geospatial distribution of risk factors for enteric diseases and malnutrition in order to extrapolate
the findings of other studies involving our population. The multi-center network will also enable
the standardization of resources, data and samples on a single platform for research, and use
of new technologies, such as those to identify and characterize host biomarkers and intestinal
microbiome related to nutritional status, disease susceptibility, and development. Defining risk
factors and biomarkers of malnutrition may eventually be developed and evaluated to build
optimal strategies for intervention and surveillance, initially at the MAL_ED network sites.
In brief, the MAL_ED network has three principal activities:

Activity 1. To establish administrative structure to develop and manage a multi-center
network in developing countries to study malnutrition and enteric infections.

Activity 2.
o
To conduct longitudinal epidemiological study, prospective and case-control to
identify factors associated with risk for enteric infections, chronic diarrhea,
malnutrition, as well as impairment of bowel function, and response to vaccine
development.
o
To collect the phenotypic and environmental data centers, enabling integration of
future studies from the expansion of collaboration in research and research
activities translated and designed to reduce infant mortality.
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
Activity 3. To develop and use temporal and geospatial models, using data from
secondary sources, like this project, to estimate the distribution of the malnutrition and
enteric infection burden, as well as the benefits attributed to various interventions.
Regarding the INCT-IBISAB, we propose that infection (and co-infection) with certain
pathogens can lead to malnutrition, due to the process of intestinal inflammation and/or altering
the intestinal barrier and its absorptive function. Thus, we test associations between specific
pathogens and malnutrition. Our study aims are to quantify these associations and thus shed
light on the following issues: (1) What organisms and mixed infections are more relevant to the
deficits in growth and development? (2) At what age in children, such infections cause more
specific impact on growth and development?
International Program of Cooperation between UFC & University of Virginia
The Clinical Research Unit (UPC) has its origin attached to the Pharmacology postgraduate course in the UFC and the international research consortium in infectious diseases
between the UFC and the University of Virginia, Charlottesville, Va, USA. In 1988, two
professors of the Pharmacology post-graduate course felt the need to establish a research core
in biomedicine at the Faculty of Medicine, focusing on research lines related to diseases that
affect directly the population of northeastern Brazil, such as infectious diseases, malnutrition,
diarrheal diseases, hypertension, diabetes, gastritis, peptic ulcers and gastric cancer. UPC
develops its research activities based on laboratory quality standards and in compliance with
ethical guidelines of national and international standard. After several training sessions and
frequent audits performed by national and international foresight such as from the Brazilian
Ministry of Health, World Health Organization, the US National Institutes of Health and the
Howard Hughes Medical Institute, the UPC acquired in 2009 the concept of global ethics in
biomedical research.
Sharing goals and objectives with IBISAB, UPC has worked hard during the biannual of
2009-2010, actively participating in the formation of two international networks, as well as the
development and creation of new postgraduate courses, as the newly created DINTER PhD
course (Inter-institutional PhD course in Morphological Sciences between UFC and UFRJ), not
neglecting the scientific productivity and training of human resources at local, national and
international levels. In this period we trained and formed four junior researchers (medical
graduate students), a Pharmacology Master Degree student and a post-doc. For details see the
website: www.upcibimed.ufc.br.
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4. Major Technical Scientific Results of INCT-IBISAB.
Patents
Listed below are the patents with records already available and are in negotiation with
the UFC rector for full import, as our original proposal. Some of these were developed within
the scientific and technological collaboration between the Federal University of Ceara & the
University of Virginia, VA, USA.
US Patent # 5,561,111
Stable Glutamine Derivatives for Oral and Intravenous Rehydration and Nutrition
Therapy (awarded the Henderson UVa. Inventor-of-the-year award).
US Patent # 6,649,746
Biological production of stable Glutamine, Poly-glutamine Derivatives in Transgenic
Organisms and Their Use for Therapeutic Purposes. Issued November 18, 2003
US Patent # 5,639,750
Method of treating C. difficile and Cholera. Issued June 17, 1997.
US Patents # 5,436, 239 and # 5,929,095
Method of treating C. difficile and Cholera. Issued July 25, 1995 and July 27, 1999.
US Patent # 4.820.714
Use of phospholipase inhibitors in the treatment of Clostridium difficile diarrhea. Issue
1994.
Biomarkers and Bioproducts Developed and / or Validated
Tables 1 to 8 showed below describe the bio-markers developed and / or validated in
the INCT-IBISAB and already available for use as DNA probes for the diagnosis of intestinal
infection, determination of virulence factors and biological studies, according to the intrinsic
goals INCT-IBISAB or those networks, RECODISA and MAL_ED which were previously
described.
Among these bio-markers, it is worth highlighting: (a) dosage of lactoferrin for
enteropathy and intestinal inflammation (Sean et al. Gastroenterology, 2010, In Press), and (b)
the test of lactulose: mannitol to determine the absorptive area, permeability and intestinal
damage (Lee et al. Gastroenteol Ped J Nutr 50: 309, 2009). Furthermore, the INCT-IBISAB
participated in the context of the international RECODISA, the development of two new bioproducts, HLZ and hG-CSF produced in milk of transgenic goats, which are currently
undergoing tests in vitro and in vivo for evaluation of their biological effects, toxicity and
effectiveness in preventive and curative therapy of diarrhea caused by bacterial agents.
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Scientific publications from INCT-IBISAB according to the thematic originally described
Thematic I
Diarrheal diseases malnutrition and oral health
Participants: Aldo AM Lima, Reinaldo B Oriá, Gerly AC Brito, Alberto M Soares, Antônio AC
Ribeiro, Helíada Chaves, Helena SA Monteiro, Júlio C Goes, Marcelo G Oliveira, Maria Cecília
F Azoubel, Renata C Leitão, Vilma Lima.
The thematic aimed at developing the following goals, for which we have reached the following
results during the year 2009:
Goals:

To determine the diarrhea etiology afflicting the semiarid children, develop molecular
methods for rapid and sensitive etiological diagnosis of the main causes of child
diarrhea and thus the bio-product development. Development of nutritional bio-products
through the interaction between the Institute and enterprises, represented by Nutrasol,
biotechnology and industry.

Investigate the production of PTX3 in human intestinal cell lines (Caco-2, T-84) and rat
intestinal cell lines (IEC-6) in vitro following exposure to bacterial toxins involved in
diarrheal diseases.

Evaluate the role of PTX 3 as an inflammation and severity marker of diarrheal diseases

Develop stable bio-products and glutamine and arginine derivates to be used as
nutritional supplementation, oral hydration and parental nutrition. These bio-products will
be developed through patents obtained by the Institute and applied to projects of the
UPC (Clinical Research Unit)

Study the modulation effect via adenosine/inosine in human intestinal cell lines (Caco-2,
T-84) and rat intestinal cell lines (IEC-6) in vitro following exposure to bacterial toxins
and in vivo models

Develop novel phospolipase A1 inhibitors for treatment of inflammatory diarrhea and
malnutrition

Study the modulation effect via adenosine/inosine in human intestinal cell lines (Caco-2,
T-84) and rat intestinal cell lines (IEC-6) in vitro following exposure to bacterial toxins
and in vivo models

Modulation of the Adenosine/inosine via as a novel therapeutic approach for diarrheal
diseases.

Evaluate the biocompatibility e biofunctionality of new biomimetic substrate complexes,
developed by the Universidade Federal do Ceara and the University of Campinas
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(UNICAMP), to be applied in tissue and bone Repair, and in the experimental
periodontal disease and in humans with chronic periodontitis.
i.
Pinto, N.B. ; Morais, T.C. ; Carvalho, K.M.B. ; Silva, C.R. ; Andrade, G.M. ; Brito, G.A.C. ;
Veras, M.L. ; Pessoa, O.D.L. ; Santos, F.A. Topical anti-inflammatory potential of
Physalin E from Physalis angulata on experimental dermatitis in mice. Phytomedicine
(Stuttgart), 10: 1016, 2010.
ii.
Sousa, Clauber M.; HAVT, Alexandre; Santos, Cláudia F.; Arnaud-Batista, F.J.; Cunha,
Karina M.A.; Cerqueira, João Batista G.; Fonteles, Manassés C.; Nascimento, Nilberto
R.F. The relaxation induced by uroguanylin and the expression of natriuretic peptide
receptors in human corpora cavernosa. The Journal of Sexual Medicine (Print), p. 1-10,
2010.
iii. Evangelista, Inez Liberato; Costa Martins, Alice Maria; Falcão Nascimento, Nilberto Robson;
HAVT, Alexandre; Azul Monteiro Evangelista, Janaina Serra; Sá de Norões, Terentia
Batista; TOYAMA, Marcos Hikari; Diz-Filho, Eduardo Brito; de Oliveira Toyama, Daniela;
FONTELES, Manassés C. Renal and cardiovascular effects of Bothrops marajoensis
venom and phospholipase A2. Toxicon (Oxford), v. 55, p. 1061-1070, 2010.
iv.
Gomes, Antoniella S; Gadelha, Gemima G; Lima, Samara J; Garcia, Joyce A; Medeiros,
Jand Venes R; Havt, Alexandre; LIMA, Aldo Ângelo Moreira; RIBEIRO, Ronaldo
Albuquerque; Cunha, Fernando Q.; Souza, Marcellus HLP. Gastro-protective effect of
heme-oxygenase 1/biliverdin/co pathway in ethanol-induced gastric damage in mice..
European Journal of Pharmacology, v. 1, p. 1-1, 2010.
v. de Freitas, MR ; de Castro Brito, GA ; de Carvalho, JV ; Gomes, RM ; Barreto Martins, MJ ;
Ribeiro, RA. Light microscopy study of cisplatin-induced ototoxicity in rats. Journal of
Laryngology and Otology, v. 123, p. 590, 2009.
vi.
Medeiros, J.V.R ; Bezerra, V.H ; Gomes, A.S ; Barbosa, A.L.R ; Lima-Junior, R.C.P ;
Soares, P. M.G ; Brito, G.A.C ; Ribeiro, R.A ; CUNHA, F.Q. ; Souza, MHLP. Hydrogen
sulphide prevents ethanol-induced gastric damage in mice: role of K+ATP channels and
capsaicin-sensitive primary afferent neurons. Journal of Pharmacology and Experimental
Therapeutics 152: 801, 2009.
vii. Freitas, MR; Silva, VC; Brito, GAC ; Carvalho Junior, JV; Gomes Junior, RM; Ribeiro, RA.
Distortion-product otoacoustic emissions and auditory brainstem responses sensitivity
assessment
in
cisplatin-induced
ototoxicity
in
rats.
Brazilian
Journal
of
Otorhinolaryngology v. 75, p. 476-484, 2009
viii. Moore SR, Lima NL, Soares AM, Oria RB, Pinkerton RC, Barrett LJ, Guerrant RL, Lima
AAM. Effects of prolonged episodes of acute diarrhea on childhood growth and risk of
14
persistent diarrhea in northeast Brazil: a community birth cohort study. Gastroenterology
000:000-000, 2010 (In Press).
ix. Quetz JS, Lima IFN, Binda AH, Carvalho EB, Lima NL, Soares AM, Mota RMS, Guerrant RL,
Lima AAM. Campylobacter jejuni and Campylobacter coli in children from communities in
Northeastern Brazil: molecular detection, intestinal inflammation and nutritional status.
Diagn Microbiol Infect Dis 00:000-000, 2010 (In Press).
x.
Ladd FV, Ladd AA, Ribeiro AA, Costa SB, Coutinho BP, Feitosa GA, de Andrade GM,
Maurício de Castro-Costa C, Magalhães CE, Castro IC, Oliveira BB, Guerrant RL, Lima
AA, Oriá RB. Zinc and glutamine improve brain development in suckling mice subjected
to early postnatal malnutrition. Nutrition. 2010 Apr 3.
xi. Oriá RB, Patrick PD, Oriá MO, Lorntz B, Thompson MR, Azevedo OG, Lobo RN, Pinkerton
RF, Guerrant RL, Lima AA. ApoE polymorphisms and diarrheal outcomes in Brazilian
shanty town children. Braz J Med Biol Res. 2010 Feb 5.
xii. LIMA, A.Â.M., Soares, AM, MOTA, RM, Maciel BL, BARRET, L.J., Fitzgerald RP, BLANER,
W. S., GUERRANT, R.L. Effects of Vitamin A Supplementation on intestinal barrier
function, growth, total parasitic and specific Giardia spp infections in Brazilian children: a
prospective randomized, double-blind, placebo-controlled trial. Journal of Pediatric
Gastroenterology and Nutrition. , v.50, p.309 - 315, 2010.
xiii. Coopeland C, Beers BB, Thompson MR, Fitzgerald RP, BARRETT, L. J., Alencar S, LIMA,
A. Â. M., GUERRANT, R.L. Faecal contamination of drinking water in a Brazilian shanty
town: importance of household storage and new human faecal. Journal of Water and
Health. v.7, p.324 - 331, 2009.
xiv. FONTELES, M.C., HAVT, A, Prata, PH, MONTEIRO, HS, LIMA, A.Â.M., Jorge, AR,
Santos, CF, GREENBERG, R.N., Nascimento NR. High-salt intake primes the rat kidney
to respond to a subthreshoul uroguanylin dose during ex vivo renal perfusion. Regulatory
Peptides. v.27, p.6 - 13, 2009.
xv. FAÇANHA, MC, Gondim AM, Pinheiro VG, Peloquin CA, GUERRANT, RL, LIMA, A.Â.M.
Intestinal barrier function and serum concentrations of rifampin, isoniazid and
pyrazinamide in patients with pulmonary tuberculosis. The Brazilian Journal of Infectious
Diseases. , v.13, p.210 - 217, 2009.
xvi. ORIÁ, RB, COSTA, CMC, LIMA, A.Â.M., PATRICK, PD, GUERRANT, R.L. Semantic
fluency: a sensitive marker for cognitive impairment in children with heavy diarrhea
burdens?. Medical Hypotheses. , v.73, p.682 - 686, 2009.
xvii. Barroso, EC, PINHEIRO, VG, FAÇANHA, MC, Carvalho, MR, Moura, ME, CAMPELO,
C.L., PELOQUIN, CA, GUERRANT, R.L., LIMA, A.Â.M. Serum concentrations of
rifampin, isoniazid and intestinal absorption, permeability in patients with multidrug
15
resistant tuberculosis. The American Journal of Tropical Medicine and Hygiene. , v.81,
p.322 - 329, 2009.
xviii. Shamir ER, Warthan M, Brown SP, Nataro JP, Guerrant RL, Hoffman PS. Nitazoxanide
inhibits biofilm production and hemagglutination by enteroaggregative Escherichia coli
strains by blocking assembly of AafA fimbriae. Antimicrob Agents Chemother. 2010
Apr;54(4):1526-33.
xix.
Pawlowski SW, Archbald-Pannone L, Carman RJ, Alcantara-Warren C, Lyerly D,
Genheimer CW, Gerding DN, Guerrant RL. Elevated levels of intestinal inflammation in
Clostridium difficile infection associated with fluoroquinolone-resistant C. difficile. J Hosp
Infect. 2009 Oct;73(2):185-7.
xx.
Mintz ED, Guerrant RL. A lion in our village--the unconscionable tragedy of cholera in
Africa. N Engl J Med. 2009 Mar 12;360(11):1060-3.
xxi. MARTINS, A. M. C. ; BARBOSA, P. S. F. ; SOUSA, D. F. ; ALVES, C. D. ; MENEZES, D.
B. ; Lima C ; FONTELES, M. C. ; LOPES-FERREIRA, M. ; MONTEIRO, H. S. A. .
Antivenom action on the renal effects induced by Thalassophryne nattereri venom.. The
Journal of Venomous Animals and Toxins Including Tropical Diseases (Online), v. 15, p.
125-135, 2009.
xxii. MARTINS, R. D. ; ALVES, R. S. ; MARTINS, A.M.C. ; BARBOSA, P. S. F. ;
EVANGELISTA, J. S. A. M. ; Evangelista, J. J. F. ; Ximenes, R. M. ; TOYAMA, M. H. ;
TOYAMA, D. O. ; Souza, A.J.F. ; Orts, D.J.B. ; MARANGONI, S. ; MENESES, D.B. ;
FONTELES, M. C. ; MONTEIRO, H. S. A. . Purification, characterization and biological
effects of phospholipase A2 from sea anemone Bunodosoma caissarum. Toxicon
(Oxford), v. 54, p. 413-420, 2009.
Books
Bases Farmacológicas e Usos Clínicos dos Antimicrobianos. Aldo A. M. Lima Editor, 1ª.
Edição, 2010. (in development).
Book chapters
LIMA, A.A.M.; GUERRANT, R.L. Cryptosporidiosis. In: Goldman; Ausiello. (Org.). Cecil
Textbook of Medicine. 23 ed.: 2010, p. 0000-0000. In Press
CARVALHO, EM; LIMA, A.Â.M. Schistosomiasis. In: Goldman; Ausiello. (Org.). Cecil Texbook
of Medicine. 23 ed.: 2010, p. 0000-0000. In Press
LIMA, A.Â.M.; SAMIE, AMIDOU; GUERRANT, R.L. Cryptosporidiosis. In: Richard L. Guerrant;
David H. Walker; Peter F. Weller. (Org.). Tropical Infectious Diseases. 3 ed.: 2010, v. 2, p.
0000-0000. In Press
16
LIMA, A.Â.M.; GUERRANT, R.L. Inflammatory Enteritis. In: Gerald L. Mandell; John E. Bennett;
Raphael Dolin. (Org.). Principles and Practice of Infectious Diseases. 7th Edition: 2010, v. 2, p.
1389-1398.
LIMA AAM, GUERRANT RL. “Cholera” in the Oxford Textbook of Medicine, 5th Edition. Edited
by Warrell DA, Cox TM, Firth JD, Benz Jr. EJ. 2010, p.754-759.
ORIÁ, RB, LIMA, AAM, GUERRANT, RL. DALYs and diarrhea In: Handbook of disease
burdens and quality of life measures, Springer Edited by Victor R. Preedy and Ronald Ross
Watson, 1st edition, 2010.
Thematic II
The second thematic proposed to screen and identify markers for prevention and treatment of
gastric cancer and ulcers, for which we give the following results for the year 2009/2010:
Aims:

Identify a typical cytokine profile associated with distal gastric carcinoma that will direct
our investigation to possible genetic mutations that might be responsible for the
increased risk to develop the disease

Pre-clinical studies of compounds isolated from medicinal plants from the Northeast in
experimental models of gastric ulcer and inflammatory bowel diseases.

Identify a specific cytokine profile that allows us to characterize individuals at risk of
developing gastric cancer and that way, guiding our investigation of genetic mutations
associated with the disease

To establish the efficacy of diterpenos alfa, beta, amirine, centipedic acid and lactone
from hawtriwaic acid in experimental models of gastric ulcer and inflammatory bowel
diseases.
QUEIROZ DMM, OLIVEIRA AG, SARAIVA IEB, ROCHA GA, ROCHA AMC, SANNA MGP,
GUERRA JB, DANI R, FERRARI MLA, CASTRO LPF. Immune response and gene
polymorphism profiles in Crohn's disease and ulcerative colitis. Inflammatory Bowel Diseases.
2009: 353-358.
QUEIROZ DM, SARAIVA IE, ROCHA GA, ROCHA AM, GOMES LI, MELO FF, BITTENCOURT
PF. IL2-330G polymorphic allele is associated with decreased risk of Helicobacter pylori
infection in adulthood. Microbes Infect. 2009; 1:80-7.
OLEASTRO M, CORDEIRO R, YAMAOKA Y, QUEIROZ D, MÉGRAUD F, MONTEIRO L,
MÉNARD A. Disease association with two Helicobacter pylori duplicate outer membrane protein
genes, homB and homA. Gut Pathog. 2009 Jun 22;1(1):12
17
OLEASTRO M, CORDEIRO R, MÉNARD A, YAMAOKA Y, QUEIROZ D, MÉGRAUD F,
MONTEIRO L. Allelic diversity and phulogeny of homB, a novel co-virulence marker of
Helicobacter pylori. BMC Microbiology, 2009, 9:248. doi: 10.1186/1471-2180-9-248
QUEIROZ DMM, CUNHA RPA, SARAIVA IB, ROCHA AMC. Helicobacter pylori virulence
factors as tools to study human migrations. Toxicon DOI: 10.1016/j.toxicon.2010.01.018
- Protective effect of anacardic acids from cashew (Anacardium occidentale) on ethanolinduced gastric damage in mice. Morais TC, Pinto NB, Carvalho KM, Rios JB, Ricardo NM,
Trevisan MT, Rao VS, Santos FA. Chem Biol Interact. 2010 Jan 5;183(1):264-9.
- Betulinic acid, a natural pentacyclic triterpenoid, prevents abdominal fat accumulation in mice
fed a high-fat diet. de Melo CL, Queiroz MG, Arruda Filho AC, Rodrigues AM, de Sousa DF,
Almeida JG, Pessoa OD, Silveira ER, Menezes DB, Melo TS, Santos FA, Rao VS. J Agric Food
Chem. 2009 Oct 14;57(19):8776-81
- Gastroprotective effect of lupeol on ethanol-induced gastric damage and the underlying
mechanism. Lira SR, Rao VS, Carvalho AC, Guedes MM, de Morais TC, de Souza AL,
Trevisan MT, Lima AF, Chaves MH, Santos FA. Inflammopharmacology. 2009 Aug;17(4):2218.
- In vitro and in vivo anti-Helicobacter pylori activity of Calophyllum brasiliense Camb. Souza
Mdo C, Beserra AM, Martins DC, Real VV, Santos RA, Rao VS, Silva RM, Martins DT. J
Ethnopharmacol. 2009 Jun 25;123(3):452-8.
Book chapter
ROCHA AMC, ROCHA GA, SOARES TF, QUEIROZ DMM. Investigação Laboratorial do
paciente com infecção por Helicobacter pylori. In: Erichsen E.S. (org). Medicina Laboratorial
para o Clínico. 1ª ED. Belo Horiozonte. Coopmed. 2009. pág 15-160
Brasil NGPS ; SILVEIRA, E.R.; CAVALCANTI, B.; Ferreira JRO ; SANTOS, F.A.; Rao VS;
COSTA-LOTUFO, L.; MORAES, M.; PESSOA, C. Chemistry and Pharmacology of Copaifera
langsdorffii Desf.. In: A.s.Awaad; J.N.Govil.; V.K.Singh. (Org.). Recent Progress in Medicinal
Plants. 1 ed. New Delhi: Studium Press, 2009, v. 27, p. 241-266
Thematic III
Autonomic Nervous System and tropical neurological diseases
Participants: Carlos M.C. Costa, Francisco A.A. Gondim, Armênio A Santos, Francisco H. Rola,
Geanne M.A. Cunha, Gisele R. Oliveira, Otoni C. Vale, Reinaldo B. Oriá
The third thematic proposed to develop 11 goals for which we reached during the year 2009 the
following results:
18
Aims:

Study of the cycle of malnutrition and infection by Cryptosporidium parvum in the
cognitive development of suckling mice

Study of the prevalence and diagnostic of neurologic diseases in the SemiArid

Research Autonomic deficiencies and nutritional modifications post-lesion of the spinal
cord in awaken mice.

Explore the impact of early malnutrition in Cryptosporidiosis in the cognitive
development in inbred BALB-C mice line.

Electrophysiological and semiological investigation of the Autonomic Nervous System
(ANS).

Correlation between the clinical manifestations and findings of the ANS examination.

Follow up of studied patients to establish prognosis, taking in account possible
cardiovascular manifestations and patient survival.

Study the chronic effect of cervical CT-scan in the gastrointestinal motility of awaken
mice.

Study the nutritional modifications in the chronic phase post-cervical and upper thorax
CT-scan.

Study whether bladder distention can promote inhibition of the digestive motility in
awaken rats post-CT-scan discharge.

Establish whether the visceral distention in animals post-CT scan discharge can
promote digestive motility inhibition without parallel changes in hemodynamic
parameters.
i. GOMES, K.N.; CÂMARA, C.C.; ARAÚJO, C.H.O.; BESSA, D.T.; LIMA, N.A.; HIRATA, F.C.C.;
COSTA, S.B.C.; TEIXEIRA-SANTOS, T.J.; ROCHA, N.M.F.L.; NORÕES, L.A.; DE CASTROCOSTA, C.M. Estudo comparativo experimental do adjuvante completo de Freund e formalina
inoculados
na
articulação
temporomandibular
de
ratos
e
consequentes
alterações
comportamentais por possível dor orofacial. Revista Dor, v. 10, p. 141-149, 2009.
ii) DE CASTRO-COSTA, C.M.; ARAÚJO, A.Q.C.; CÂMARA, C.C.; FERREIRA, A.S.;
TEIXEIRA-SANTOS, T.J.; COSTA, S.B.C.; ALCÂNTARA, R.N.M.; TAYLOR, G.P. Pain in
tropical spastic paraparesis/HTLV-I associated myelopathy patients. Arquivos de NeuroPsiquiatria v. 67, p. 866-870, 2009.
iii) DE CASTRO-COSTA, C.M. Dor no Câncer: Tópicos Relevantes. Âmbito Hospitalar, v. 5, p.
80-83, 2009.
iv) DE CASTRO-COSTA, C.M. Dor Neuropática Novos Conceitos, Expressão Clínica e
Desafios Terapêuticos. Prática Hospitalar v. 1, p. 109-110, 2009.
19
v) BATISTA, L.M.; BATISTA, I.M.; ALMEIDA, J.P.; CARVALHO, C.H.; COSTA, S.B.C.; DE
CASTRO-COSTA, C.M. Preemptive Analgesic Effect of Lidocaine in a Chronic Neuropathic
Pain Model. Arquivos de Neuro-Psiquiatria v. 67, p. 1088-1092, 2009.
vi) GONDIM, F.A.A.; THOMAS, F.P.; OLIVEIRA, G.R.; PIMENTEL, L.H.C.; BASTOS, B.P.R.;
DE CASTRO-COSTA, C.M. On the spectrum of leprosy neuropathies: multifocal inflammatory
neuropathy heralding leprosy relapse. Neuromuscular Disorders, v. 19, p. 711-713, 2009.
vii) ORIÁ, R.B.; DE CASTRO-COSTA, C.M.; LIMA, A.A.M.; PATRICK, P.D.; GUERRANT, R.L.
Semantic fluency: a sensitive marker for cognitive impairment in children with heavy diarrhea
burdens? Medical Hypotheses, v. 73, p. 682-686, 2009.
i. Castro-Silva, C.; BRUIN, V. M. S.; CUNHA, GMA; Nunes, D.M.; Medeiros, C.A.M.; BRUIN, P.
F. C. Melatonin improves sleep and reduces nitrite in the exhaled breath condensate in cystic
fibrosis a randomized, double-blind placebo-controlled study. Journal of Pineal Research, v. 48,
p. 65-71, 2010.
ii. RODRIGUES, P.A.; MORAES, S.M.; SOUZA, C.M.; SILVA, A.R.; ANDRADE, GM; Silva,
M.G.; RAO, Vietla Satyanaraiana; SANTOS, F.A. Gastroprotective effect of barbatusin and 3beta-hydroxy-3-deoxibarbatusin, quinonoid diterpenes isolated from Plectranthus grandis, in
ethanol-induced gastric lesions in mice. Journal of Ethnopharmacology, v. 127, p. 725-730,
2010.
iii. PINTO, N.B.; MORAES, T.C.; CARVALHO, K.M.; SILVA, C.R.; ANDRADE, GM; BRITO,
G.A.C.; PESSOA, O.D.L.; SANTOS, F.A. Topical anti-inflammatory potential of Physalin E from
Physalis angulata on experimental dermatitis in mice. Phytomedicine (Stuttgart), v. 8, p. 00-01,
2010
iv. NOBRE JUNIOR, H.V.; OLIVEIRA, RA; MAIA, F.D.; NOGUEIRA, M.A.S.; MORAES, M.O.;
BANDEIRA, M.A.M; CUNHA, GMA; VIANA, GSB. Neuroprotective Effects of chalcones from
Myracrodruon urundeuva on 6-Hydroxydopamine-Induced cytotoxicity in rat mesencephalic
cells. Neurochemical Research, v. 34, p. 1066-1075, 2009.
v. LEAL, LKAM; CANUTO, K.M.; Costa, K.C.; NOBRE JÚNIOR, HV; VASCONCELOS, S.M.M.;
SILVEIRA, ER; Ferreira, M.V.; FONTENELE, J.B.; ANDRADE, GM; VIANA, GSB. Effects of
Amburoside A and Isokaempferide, Polyphenols from Amburana cearensis on rodent
inflammatory processes and myeloperoxidase activity in human neutrophils activity in human
neutrophilis. Basic & Clinical Pharmacology & Toxicology, v. 104, p. 198-205, 2009.
vi. BRUNO, R; MARQUES, T; BATISTA, T; LIMA, J; DEARRUDA, K; LIMA, P; SANTOS, N;
CUNHA, GM; VITOR, H; VIANA, GSB. Pentoxifylline treatment improves neurological and
neurochemical deficits in rats subjected to transient brain ischemia. Brain Research, v. 1260, p.
55-64, 2009.
vii. CANAS, P.; PORCIUNCULA, L. O.; CUNHA, GM; SILVA, C.G.; MACHADO, N.J.;
OLIVEIRA, J.M.; OLIVEIRA, C. R.; CUNHA, R.A. Adenosine A2A receptor blockade prevents
20
synaptotoxicity and memory dysfunction caused by beta-amyloid peptides via p38 mitogenactivated protein kinase pathway. The Journal of Neuroscience, v. 29, p. 14741-14751, 2009
i. PEIXOTO Jr. AA; TELES BC; BRASIL EF; Santos AA; Oliveira GR; Ribeiro RA; Rola FH;
Gondim FAA. Vincristine delays Gastric Emptying and Gastrointestinal Transit of Liquid in
Awake Rats. Brazilian Journal of Medical and Biological Research, v. 42, p. 567-573, 2009.
ii. Oliveira GR; Gondim FAA; Hogan E; Rola FH. Movement-Induced heart rate (HR) changes in
epileptic and non-epileptic seizures. Arquivos de Neuro-Psiquiatria, v. 67, p. 789-791, 2009
iii. Gondim FAA; Oliveira GR; Thomas FP. Upper gastrointestinal motility changes following
spinal cord injury. Neurogastroenterology and Motility, v. 22, p. 2-6, 2010
iv. Gondim FAA; Thomas FP; OLIVEIRA G; Pimentel LH; Costa CM de C. On the spectrum of
leprosy
neuropathies:
multifocal
inflammatory
neuropathy
heralding
leprosy
relapse.
Neuromuscular Disorders, v. 19, p. 711-713, 2009.
v. TAUNAY T; Oliveira GR; Gondim FAA. Neuroanálise: considerações sobre as bases
neurobiológicas das doenças psiquiátricas. Rheumatology (Oxford), v. 2, p. 5-10, 2009
Books:
i. ALVES-NETO, O. (Org.); DE CASTRO-COSTA, C.M. (Org.); SIQUEIRA, J.T.T. (Org.);
TEIXEIRA, M. J. (Org.). Dor: Princípios e Prática. 1. ed. Porto Alegre-RS: Artmed Editora,
2009. v. 1. 1438 p.
i. Gondim FAA; TAUNAY T; Oliveira GR; Messias EL; Costa CM de C; Rola FH; Costa HA.
Neuropsicofisiologia. 1. ed. Fortaleza: Gráfica Batista, 2009. v. 1. 257 p.
Book chapters:
i. DE CASTRO-COSTA, C.M.; TEIXEIRA-SANTOS, T.J.; COSTA, S.B.C. Modelos animais e
laboratoriais de dor. In: NETO, O.A.; DE CASTRO-COSTA, C.M.; SIQUEIRA, J.T.T.;
TEIXEIRA, M.J. (Org.). Dor: Princípios e Prática. 1 ed. Porto Alegre-RS: Artmed Editora, 2009,
v. 1, p. 305-312.
ii. DE CASTRO-COSTA, C.M. Dor Neuropática. In: NETO, O.A.; DE CASTRO-COSTA, C.M.;
SIQUEIRA, J.T.T.; TEIXEIRA, M.J. (Org.). Dor: Princípios e Prática. 1 ed. Porto Alegre-RS:
Artmed Editora, 2009, v. 1, p. 495-509.
iii. VALL, J.; DE CASTRO-COSTA, C.M. Dor em Lesão Medular. In: NETO, O.A.; DE CASTROCOSTA, C.M.; SIQUEIRA, J.T.T.; TEIXEIRA, M.J. (Org.). Dor - Princípios e Prática. 1 ed. Porto
Alegre-RS: Artmed Editora, 2009, v. 1, p. 838-846.
i. SILVA JUNIOR JTF; MONTEIRO IGL; HOLANDA CTD; Gondim FAA. Neurological
Complications of Antiangiogenic Therapy. In: Malay Chatterjee; Ajay Rana; Basabi Rana.
(Org.). Anti-Angiogenesis-Drug Discovery and Development. Sharjah: Bentham Science
Publishers, 2010.
21
Thematic IV
The fourth thematic (Inflammation and gastrointestinal motility) included five goals from which
we give the following achievements during the year 2009:
Aims:
•
Modulation of the inflammatory response, as threrapeutic strategy to revert the digestive
motility alterations, associated with chemotherapy-induced mucositis
•
Investigate the contractility of the smooth muscle of the gastric fundus and duodenum in
vitro induced by KCl, serotonine, carbacol or electrical stimulation from rats or mice pre-treated
with 5-FU or irinotecan.
•
Study the pharmacological characteristics of terpenos (alfa and beta-pireno as well as
others), found in plant oils from the Northeastern Semiarid, applied to the gastrointesinal tract.
•
Investigate whether self-antibodies against enteric ionic channels are present and/or
whether are useful for diagnosis of patients with DNMGI. Investigate if the self-antibodies (if
present) are pathogenic
•
Contribute to elucidate the expression profile of ionic channels in the enteric
neuromuscular layers, including CIC
1. Barbosa, A. L. R. ; Pinheiro, C. A. ; Oliveira, G. J. ; Moraes, M. O. ; RIBEIRO, R. A. ; VALE,
M. L. ; Souza, M. H. L. P. . Tumor bearing decreases systemic acute inflammation in rats role
of mast cell degranulation. Inflammation Research, p. 1-2, 2009.
2. Gomes, Antoniella Souza ; Lemos, Henrique Paula ; Medeiros, Jand Venes Rolim ; Cunha,
Fernando Queiroz ; Souza, Marcellus Henrique Loiola Ponte . Lipopolysaccharide from
Escherichia coli prevents indomethacin-induced gastric damage in rats: role of non-protein
sulfhydryl groups and leukocyte adherence. Inflammation Research, p. 10-11, 2009.
3. Medeiros, J. V. R ; Bezerra, V. H ; Gomes, A. S ; Barbosa, A. L. R ; Lima-Junior, R. C. P ;
Soares, P. M. G ; Brito, G. A. C ; Ribeiro, R. A ; CUNHA, F. Q. ; Souza, M.HL P. Hydrogen
sulphide prevents ethanol-induced gastric damage in mice: role of katp channels and capsaicinsensitive primary afferent neurons. Journal of Pharmacology and Experimental Therapeutics, p.
2, 2009.
4. Murad-Regadas, Sthela M. ; Regadas, Francisco Sérgio P. ; Rodrigues, Lusmar V. ; Oliveira,
Leticia ; Barreto, Rosilma G. L. ; Souza, M. H. L. P. ; SILVA, F. R. S. . Types of pelvic floor
dysfunctions in nulliparous, vaginal delivery, and cesarean section female patients with
obstructed defecation syndrome identified by echodefecography. International Journal of
Colorectal Disease , v. 24, p. 1227-1232, 2009.
22
5. Souza, M. A. N. ; Souza, M. H. L. P. ; Palheta, R. C. ; Cruz, P. R. M. ; MEDEIROS, B. A. ;
ROLA, F. H. ; Magalhaes, P. J. C. ; TRONCON, L. E. A. ; SANTOS, A. A. . Evaluation of
gastrointestinal motility in awake rats: a learning exercise for undergraduate biomedical
students. Advances in Physiology Education, v. 33, p. 343-348, 2009.
6. Souza Filho, M.V.P. ; Loiola, R.T. ; Rocha, E.L. ; Simão, A.F.L. ; Gomes, A.S. ; Souza, M. H.
L. P. ; RIBEIRO, R. A. . Hind limb ischemic preconditioning induces an anti-inflammatory
response by remote organs in rats. Brazilian Journal of Medical and Biological Research, v. 42,
p. 921-929, 2009.
1.
Ferreira W ; Barbosa R ; Nascimento T ; Moreira. L ; Oliveira D ; SOUZA, Andrelina
Noronha Coelho de ; MAGALHÃES, PJC ; LAHLOU, S ; CARDOSO, José Henrique Leal .
Effects of 1,8-cineole on electrophysiological parameters of neurons of rat superior cervical
ganglion. Clinical and Experimental Pharmacology & Physiology, v. 36, p. 1068-73, 2009.
2. BASTOS, Vasco Pinheiro Diogenes ; BRITO, Teresinha Silva de ; BATISTA- LIMA, F. J. ;
PINHO, J. P. M. ; LAHLOU, S ; MATOS, Francico Jose de Abreu ; SANTOS, AA ;
MAGALHÃES, PJC . Inhibitory effect of 1,8-cineole on guinea pig airway challenged with
ovalbumin involves a preferential action on electromechanical coupling. Clinical and
Experimental Pharmacology & Physiology, v. 36, p. 1120-6, 2009.
3. PORTO, N. P. C. ; JUCÁ, Davi Mathews ; LAHLOU, S ; COELHO-DE-SOUZA, AN ;
DUARTE, Gloria Isolina Buentes Pinto ; MAGALHÃES, PJC . Effects of K+ channels inhibitors
on the cholinergic relaxation of the isolated aorta of adult offspring rats exposed to maternal
diabetes. Experimental and Clinical Endocrinology & Diabetes, 2009 (in press).
4. Leonhardt, V. ; LEAL-CARDOSO, JH ; LAHLOU, S ; ALBUQUERQUE, Aline Alice
Cavalcante de ; PORTO, R. S. ; CELEDONIO, N. R. ; OLIVEIRA, A. C. ; MAGALHÃES, PJC ;
DUARTE, Gloria Isolina Boentes Pinto ; COELHO-DE-SOUZA, AN . Antispasmodic effects of
the essential oil of Pterodon polygalaeflorus and its main constituent b-caryophyllene on rat
isolated ileum. Fundamental & Clinical Pharmacology, 2009 (in press).
5. Moreira-Lobo DC, Linhares-Siqueira ED, Cruz GM, Cruz JS, Carvalho-de-Souza JL, Lahlou
S, Coelho-de-Souza AN, Barbosa R, Magalhães PJ, Leal-Cardoso JH. Eugenol modifies the
excitability of rat sciatic nerve and superior cervical ganglion neurons. Neurosci Lett. 2010.
6. Lima FJ, Brito TS, Freire WB, Costa RC, Linhares MI, Sousa FC, Lahlou S, Leal-Cardoso
JH, Santos AA, Magalhães PJ. The essential oil of Eucalyptus tereticornis, and its constituents
alpha- and beta-pinene, potentiate acetylcholine-induced contractions in isolated rat trachea.
Fitoterapia. 2010 Mar 17,2010 Mar 26;472(3):220-4
i. Cristino-Filho, G. ; Leal, P.R.L. ; Lira, G.H.S. ; Souza, S.G. ; Rola, F.H. ; Oliveira, R.B. ;
Souza, M.A.N. ; Santos, A. A. . Acute intracranial hypertension increases gastric tonus in
anesthetized rats. Autonomic Neuroscience, v. 146, p. 95-101, 2009.
23
ii. Albuquerque Filho, F.L. ; Macedo, Geraldo Munguba ; Santos, A.A. ; Rodrigues, L.M. ;
Oliveira, R.B. ; Souza, M.A.N. . Stapled haemorrhoidopexy decreases transiently rectal
compliance and sensitivity threshold. Colorectal Disease, v. XXX, p. x, 2009.
iii. Bruin, Veralice Meireles Sales de ; Noronha, AC ; Souza, MAN ; Araújo, RP ; Mota, RMS ;
Bruin, PFC . Gastroesophageal reflux and obstructive sleep apnea in childhood. International
Journal of Pediatric Otorhinolaryngology, v. 73, p. 383-389, 2009.
iv. Peixoto Júnior, A.A. ; Teles, B.C.V ; Castro, E.F.B. ; Santos, A.A. ; de Oliveira, G.R. ;
Ribeiro, R.A.; Rola, F.H. ; Gondim, F.A.A. . Vincristine delays gastric emptying and
gastrointestinal transit of liquid in awake rats. Brazilian Journal of Medical and Biological
Research, v. 42, p. 567-573, 2009.
v. ARCANJO, F.P.N.; Amancio, O.M.S.; Pinto, V.P.T. Effect of a beverage fortified with
evaporated sugarcane juice on hemoglobin levels in preschool children. Pan American Journal
of Public Health v. 26, p. 350-354, 2009.
vi. PINTO, V.P.T.; Nascimento, K.S.; SAMPAIO, A.H. ; TEIXEIRA, E.H.; Teixeira, A.H.;
CAVADA, B.S.; Debray, H. Lectins from the red marine algal species Bryothamnion seaforthii
and Bryothamnion triquetrum as tools to differentiate human colon carcinoma cells. Advances
in Pharmacological Sciences, v. 2009, p. 1-7, 2009.
vii. PALHETA JÚNIOR, Raimundo Campos ; Rola, Francisco Hélio ; LIRA, Gislano Heverton
Soares de ; Gomes, Dayane Aparecida ; CARVALHO, Fernando Mota ; Elias, Lucila Leico
Kagohara ; Antunes-Rodrigues, José ; OLIVEIRA, Ricardo Brandt de ; Santos, A.A. . Atrial
stretch increases the gastric tonus of anesthetized rats. Life Sciences (1973), 2010.
Articles accepted for publication
1.GOMES, A. S. ; Gadelha, G G ; Lima S J ; GARCIA, J. A. ; Medeiros, Jand Venes Rolim ;
LIMA, A. A. M. ; Havt, A ; RIBEIRO, R. A. ; BRITO, G. A. C. ; Souza, M. H. L. P. .
Gastroprotective effect of heme-oxygenase 1/biliverdin/CO pathway in ethanol-induced gastric
damage in mice. European Journal of Pharmacology, 2010.
2. JUNQUEIRA JÚNIOR, J. ; LIMA-JÚNIOR RCP ; MEDEIROS, J. R. ; BRITO, G. A. C. ;
RIBEIRO, R. A. ; SOUZA, M. H. L. P. . Role of capsaicin sensitive primary afferent neurons and
non-protein sulphydryl groups on gastroprotective effect of amifostine against ethanol-induced
gastric damage in rats. Digestive Diseases and Sciences, 2010.
Leal-Cardoso, JH, Lahlou S, Weinreich D, Magalhães PJC. The essential oil of Croton
nepetaefolius selectively blocks histamine-augmented neuronal excitability in guinea-pig celiac
ganglion. J Pharm Pharmacol, 2010.
Book chapters:
24
SANTOS, A. A. . Motilidade gastrintestinal. In: Rui Curi; Joaquim Procópio Araújo Filho. (Org.).
Fisiologia Básica. 1 ed. Rio de Janeiro: Guanabara Koogan, 2009, v. Cap43, p. 609-624.
RIBEIRO, R. A. ; VALE, M. L. ; Souza, M H L P . ANTIINFLAMATÓRIOS NÃO HORMONAIS E
COXIBES. In: Onofre Alves Neto; Carlos Maurício de Castro Costa, José Tadeu T. de Siqueira,
Manoel Jacobsen Teixeira. (Org.). DOR PRINCÍPIOS E PRÁTICA.. Porto Alegre: Artmed,
2009, v. 1, p. 1056-1073.
Souza, MAN. Abordagem do paciente com disfagia. In: José Milton de Castro Lima. (Org.).
Gastroenterologia. Fortaleza: Edições UFC, 2009.
Article in Science Education (Physiology)
Souza, M. A. N.; Souza, M.H.L.P.; Palheta, R.C.; Cruz, P.R.M.; Medeiros, B.A.; Rola, F.H.;
Magalhães, P.J.C.; Troncon, L.E.A. ; Santos, A.A. Evaluation of gastrointestinal motility in
awake rats: a learning exercise for undergraduate biomedical students. Advances in Physiology
Education, v. 33, p. 343-348, 2009.
5. INCT-IBISAB National and International Events.
In this reporting period, the INCT-IBISAB organized and fostered two international
events: (1) V International Symposium in Biomedicine and (2) XIV Tropical Medicine &
International Collaborative Research Center, Infectious Diseases Research. These events
served as an opportunity for the encounter of different INCT-IBISAB research groups, well as
the interaction of national and international guests, allowing information exchange and the
eventual establishment of collaborative projects. The programs of these two events are
attached to the report, as below:
Summary of the 14th Tropical Medicine Research Center (TMRC) and International
Collaboration in Infectious Diseases Research (ICIDR),
Fortaleza, Ceará, March 5-6, 2010
Opened by our host, Dr. Aldo A.M. Lima, the first session focused on enteric infections
and malnutrition, with the first talk by Drs. Alexandre H. Binda and Ila F. N. Lima being on the
high prevalence and genetic heterogeneity of E. coli pathotypes infecting young children in
Northeast Brazil. This work showed that the majority of children are colonized with potentially
pathogenic genotypes of E. coli, regardless the presence of overt diarrheal symptoms. This
provoked excellent discussion about the potential importance of quantitative cultures or colony
selection or of quantitative PCR to distinguish clinically significant levels of infection.
Next, Dr. Aldo Lima provided an overview of mechanisms and assessment of intestinal
barrier disruption, first reviewing the results from an extensive trial of vitamin A, zinc, and
glutamine with 8 blinded permuted blocks with all combinations and with placebo controls.
These showed that low serum glutamine levels correlated closely with low serum arginine
25
levels, and they correlated with impaired growth and disrupted intestinal barrier function. He
also described the impressive impact of glutamine with vitamin A and zinc on improved verbal
learning in the girls, but with less apparent effects in the boys. Finally he described improved
growth (by HAZ) and WRAML learning scores after glutamine treatment, but primarily in the
ApoE4+ children, suggesting a synergistic effect of glutamine in E4 children on growth and
learning.
Then Dr. Sean Moore presented a reappraisal of diarrhea durations, showing that
“prolonged” diarrhea (ie lasting >7d) accounted for fully half of the days of diarrhea, and, like
“persistent” diarrhea (lasting >14d), prolonged diarrhea also identifies children in whom one can
predict heavy diarrhea burdens, with some similar etiologies (ie Cryptosporidium and Shigella)
as persistent diarrhea. These studies have key implications for identifying the high risk child in
need of effective interventions to avoid the risk of long-term effects of heavy diarrhea burdens.
Dr. Reinaldo B. Oria next described how zinc and glutamine protect hippocampal brain
development in a malnourished neonatal murine model, with both improved WAZ and HAZ and
cliff avoidance with zinc (with or without glutamine). He also described significant hippocampal
hypertrophic changes and neuronal swelling with malnutrition, and finally, improvement in
synaptophysin levels and brain zinc and hippocampal GABA levels with glutamine treatment.
Lastly in this session, Dr. Richard L. Guerrant described new models, host defenses and
novel therapeutic approaches for enteric infectious etiologies of malnutrition, with similar impact
on growth by cryptosporidial infections and EAEC infections in weaned C57Bl6 mice as seen in
the neonatal mice, and with enhancement of infection and of impact on growth in protein
deprived malnourished weaned mice. Furthermore, the weaned model allows assessment of
adaptive as well as innate immune responses, and initial findings with a ClyA-CP-15 Salmonella
vaccine given intranasally (in collaborations with Drs. Jim Galen at UMd and Greg Buck and
Patricio Manque at VCU) shows for the first time, greatly impaired antibody responses to a
mucosal vaccine in malnourished weaned mice, to open studies of mechanisms and
interventions for mucosal vaccine failure (or reduced efficacy) in endemic areas (as seen for
OPV and rotavirus vaccines).
Initial studies also suggest alterations in cytokine and TLR
expression with malnutrition, as well as CpG and even non-CpG ODN reduction of
cryptosporidial infections in HCT-8 human colonic intestinal cells in vitro.
The afternoon session was opened by Dr. Warren D. Johnson Jr., who had been
honored earlier for his lifelong commitment to collaborations in endemic tropical areas,
speaking about his recent work with extraordinary and heroic colleagues in Haiti following the
tragic earthquake there.
Then the second session, chaired by Drs. Johnson and Edgar Carvalho (Director of the
TMRC) focused on the “Host Immunologic and Parasite Factors in the Pathogenesis of Dermal
Leishmaniasis”, with the opening paper by Dr. Albert Schreifer addressing the role of
26
Leishmania braziliensis polymorphisms in disease outcomes and distribution. He described
impressive differences in clades of L. braziliensis in cutaneous, mucosal and disseminated
cutaneous leishmaniasis as well as in geographic areas affected in the Corte de Pedra study
field.
This was followed by Dr. Edgar M. Carvalho, Director of the TMRC, describing
protective and pathogenic immune responses in tegumentary leishmaniasis, with Th1 and
inflammatory cell predominance in cutaneous and mucosal leishmaniasis, with increased CD8
CTLs and granzyme expression in ulcer lesions, but no defect in Treg cells, and disappearance
of proliferative and cytokine responses over time after subclinical infections. Thus adaptive
immune responses correlate with pathology, not protection, the latter for which innate immunity
may be key. Then Dr. Lucas P. Carvalho described his work on the role of monocytes in the
pathogenesis of cutaneous leishmaniasis, showing that ulcers are associated with
“proinflammatory” monocytes expressing CD16 as well as CD 14 markers.
Finally in this session, Dr. Phil Scott from the University of Pennsylvania described how
lymph node hypertrophy enables the host to maximize its response to leishmaniasis. Using his
C57Bl6 murine model, he described how CCL21 chemokines stimulate CCR7 receptors on
bystander (uninfected) T cells as well as dendritic cells.
He also described the TLR9
dependence of this response and of protection, using TLR9ko mice.
Chaired by Drs. Selma M.B. Jeronimo on Saturday morning, March 6th, the 3rd session
focused on Leishmaniasis genetic determinants and diagnosis, with the opening talk by Dr.
Mary E. Wilson being on “A species discriminating diagnostic quantitative PCR for
leishmaniasis enumeration of kDNA copies in Leishmania species, strains and life stages.”
This provides a web-described tool for assessment of infection severity in humans, dogs or
sandflies, and opens new levels of epidemiologic and pathogenetic studies.
Then Dr. Selma Jeronimo described her observations over 20 years from the urban
fringe on Leishmania chagasi infection in northeastern Brazil, suggesting that the infected sand
fly vector Lutzomyialongipalpis is becoming more anthropophilic (or at least in settings of
human crowding, human reservoirs become key hosts for sandfly infection). Remarkably, she
described that up to 27% of female sandflies can be infected in endemic houses, and that some
30% of seronegative dogs in the area develop serologic evidence of infection over a 6-month
follow-up period, and similar numbers are seropositive and revert to seronegative.
Dr. Carlos E.M. Gomes did not attend to describe his work on the ethnic structure of
populations living in endemic areas for Leishmania and Mycobacterium infections in North and
Northeast Brazil, so next, Bruna L. L. Maciel then described her work on the impact of
Leishmania chagasi infection on the nutritional status and child development. Though HAZ and
WAZ were not affected (in these slightly older children with VL infections –than in the early
childhood diarrhea studies), L. chagasi infections were associated with reduced skin-fold
thickness and body mass index, as well as reduced serum retinol (and reduced vitamin A
27
stores, as assessed by binding competition assay) and with acute phase reactants, CRP and
alphaGP. She also noted that children with higher weight at birth were protected, and that
longer breastfeeding was associated with increased evolution to asymptomatic infection.
The final session on Tuberculosis and HTLV-1 infection included Dr. Lee W. Riley’s
presentation on his seminal works on a post-exposure vaccine for tuberculosis, and its
potentially important role in the tuberculosis eradication. Using his mce1 gene mutant strains,
in particular his mce1R mutant (that inactivates the negative repressor to cause
hyperexpression) that provides, after only 4 weeks of INH and PZA treatment in the Cornell
Balb/c murine model, a model of reactivation after few weeks of latency, he used a mce1A
piece to provide a vaccine that prevents reactivation. As animal studies, especially with M.
tuberculosis, are difficult, he suggested that this vaccine may soon require direct testing in
humans.
Dr. Edgar M. Carvalho then described his work on the viral and immunological factors
in clinical manifestations associated with HTLV-1 infection. While HAM/TSP may only affect
<5% of HLV-1 infected subjects, up to 40-60% of previously thought “asymptomatic” patients
have neurogenic bladder and correlated erectile dysfunction, even at younger ages of 30-40
years old. He further described associations of these urologic symptoms with increased TNFalpha and IFN-gamma (and higher proviral loads), compared to HTLV-1 infections without
these symptoms, suggesting similar immunopathogenesis to that seen with HAM/TSP.
Implications for pentoxyphylline, thalidomide and rolipram therapy to reduce TNF and/or IFN
(pentoxyphylline and rolipram reduce both; thalidomide only TNF) as well as planned clinical
trials were suggested.
Dr. Richard L. Guerrant then made his closing remarks with the four-session overview
over two days of the 14th TMRC meeting, impressively spanning the innovative and productive
outcomes of the sharing collaboration and partnerships that have clearly fostered novel work.
Work that has benefited tremendously the understanding of major endemic diseases afflicting
the poor in Northeast Brazil. The richness of discussions throughout the meeting days across
all groups was yet the best, as this Center matures into trusting open collaborations across
multiple disciplines within Brazilian and US institutions.
Close celebrations in Dr. Johnson’s
honor in Salvador and in the Dr. Lima’s V. International Symposium in Biomedicine in Fortaleza
not only took advantage of participant gathering from multiple institutions in Brazil and the US,
but also bridged further the interdisciplinary and institutional collaborations, as exemplified by
Dr. Peter Patrick, neuropsychologist from UVa and Drs Sidarta Ribeiro and Draulio de Araujo,
neurobiologist modelers and MRI neurophysicists, respectively, from UFRN, who described
neurocognitive and radiological brain assessments relevant to on going work on cognitive
development with diarrhea, enteric infections and leishmaniasis, with possible collaborations in
the future. Signed by Richard L. Guerrant, M.D.,
March 6, 2010.
28
6. Training activities and training of human resources in INCT-IBISAB
As a direct offshoot of research activity undertaken by INCT-IBISAB qualified member
team occurred as follows:
Thematic I
Doctoral Thesis
Elisabete Clara Barroso. Study of intestinal barrier function and serum concentrations of
rifampicin and isoniazid in patients with multidrug resistant tuberculosis. 2009. Doctor in
Medical Sciences - UFC. Advisor: Aldo Ângelo Moreira Lima.
Inez Liberato Evangelista. Characterization of cardiorenal and neural activities of Bothrops
marajoensis venom and its fractions. 2009. Doctor in Pharmacology - UFC. Advisor: Helena
Serra Azul Monteiro.
Master’s dissertation
Josiane da Silva Quertz. Study on Campylobacter jejuni and Campylobacter coli in children
from urban area in Fortaleza, Ceará/Brazil. 2009. Master Degree in Pharmacology - UFC.
Advisor: Aldo Ângelo Moreira Lima.
Diana Maria de Almeida Lopes. Determining the profile of patients and evaluation of treatment
of latent tuberculosis infection in candidates for the use of TNF-alpha blockers in a university
hospital (HUWC/UFC). 2010. Master Degree in Pharmacology, UFC. Advisor: Helena Serra
Azul Monteiro.
Claudênio Diógenes Alves. Study of renal effects of the venom of the snake Lachesis muta
muta. 2010. Master Degree in Pharmacology, UFC. Advisor: Helena Serra Azul Monteiro.
Antonio Rafael Coelho Jorge. Study the effects of high intake of sodium chloride orally daily on
the metabolism and renal function in rats. 2009. Master Degree in Pharmacology, UFC.
Advisor: Helena Serra Azul Monteiro.
Rafael Matos Ximenes. Evaluation of the activity of A-II cabenegrins on the biochemical,
hematological and blood pressure effects induced by the venom of Bothrops jararacussu in
rats. 2009. Master Degree in Pharmacology, UFC. Advisor: Helena Serra Azul Monteiro.
Terentia Batista de Sá Norões. Renal effects promoted by venom of Bothrops atrox and
systemic release of nitric oxide. 2009. Master Degree in Pharmacology, UFC. Advisor: Helena
Serra Azul Monteiro.
Thematic II
Doctoral Thesis
29
Marjorie
Moreira
Guedes.
Investigation
of
the
pharmacological
mechanisms
of
gastroenteroprotectors action of acid centipede, a diterpene of Egletes viscosa Less. in
experimental models. Doctor in Medical Sciences UFC. Advisor:: Vietla Satyanarayana Rao
Caroline Mourão Melo. Study of the pharmacological effect of alpha, beta-amyrin, a triterpene
mixture isolated from Protium heptaphyllum in experimental acute pancreatitis. Doctor in
Pharmacology, UFC. Advisor: Flávia Almeida Santos
Master’s dissertation
Talita Cavalcante Morais. Effect Analgesic, Anti-inflammatory and gastroprotective of anacardic
acid, isolated from Anacardium occidentale L., in Experimental Models. Master Degree in
Pharmacology, UFC. Advisor: Flávia Almeida Santos
Cícero Igor Simões Moura Silva: Genotypes cagA, and vacA alleles and tyrosine
phosphorylation sites of CagA protein of H. pylori in patients with and without family history of
gastric cancer. Master Degree in Medical Sciences - UFC, Advisor:: Lucia Libanez Bessa
Campelo Braga UFC Data da defesa: 16/12/2009.
Sérgio de Assis Batista – “Evaluation of the pattern and number of sites EPIYA CagA protein of
Helicobacter pylori and risk of gastric carcinoma and duodenal ulcer”. Master Degree in
Microbiology, ICB/UFMG Advisor: Dulciene Queiroz
Thematic III
Master’s dissertation
i. Ayrton Silva Ferreira. Pain Associated Myelopathy Virus Human T cell Lymphotropic Type-I
(HTLV-I). 2009. Master Degree in Pharmacology - UFC, Advisor: Carlos M.C Costa.
ii. Ailton Teles Fontenele Filho. Effect of psychosocial stress on brain damage induced by
cerebral ischemia. Role of adenosine A1 receptor. 2009. Master Degree in Pharmacology UFC. Advisor: Geanne M Cunha.
iii. Marta Regina Santos do Carmo. Effect of PPADS, P2 receptor antagonist, on brain damage,
behavior and memory in ischemic mice. 2009. Master Degree in Pharmacology - UFC, Advisor:
Geanne M Cunha.
Doctoral Thesis
i. Carlos Campos Câmara. Study of the analgesic, behavioral, and regenerative therapy with
gabapentin in experimental models of neuropathic pain. 2009. Doctor in Pharmacology - UFC,
Advisor: Carlos M.C Costa.
Thematic III
Doctoral Thesis
30
Raimundo Campos Palheta Júnior. Characterization of neurohumoral pathways in delayed
gastric emptying of liquids arising from right atrial mechanical distension in awake rats. 2010.
Doctor in Pharmacology – UFC. Advisor: Armenio Aguiar dos Santos.
Vasco Pinheiro Diógenes Bastos. Bronchodilator and anti-inflammatory action of 1,8-cineole in
experimental asthma in guinea pigs. Doctor in Pharmacology – UFC, 2009. Advisor: Pedro
Jorge Caldas Magalhães
Antoniella Souza Gomes. Protective effect via hemeoxigenase 1 / biliverdin / co models of
gastric lesions in mice role of soluble guanylate cyclase (SCG) and NO synthase (NOS). 2009.
Doctor in Pharmacology – UFC. Advisor: Marcellus Henrique Loiola Ponte de Souza.
Jand-Venes Rolim Medeiros. Gastroprotective and prokinetic effect of hydrogen sulfide (H2S)
in mice models, the role of afferent neurons sensitive to capsaicin receptor TRPV1 channels
and ATP dependent K + (KATP). Doctor in Pharmacology – UFC. Advisor: Marcellus Henrique
Loiola Ponte de Souza. 2009.
Master’s dissertation
José Nelson Belarmino Filho. Mucositis induced irinitecan (cpt11) - The role of leukocyteendothelial interaction. 2010. Master Degree in Pharmacology, UFC. Advisor: Marcellus
Henrique Loiola Ponte de Souza
Larisse Tavares Lucetti. Effect of pentoxifylline and dexamethasone on inflammatory response
and changes in gastrointestinal motility associated with intestinal mucositis induced by 5 fluorouracil in rats. 2009. Master Degree in Pharmacology, UFC. Advisor: Marcellus Henrique
Loiola Ponte de Souza.
Monalysa Neves Costa. Identification, isolation and physicochemical characterization of
molecular markers present in erythrocytes of sickle cell anemia. 2009. Master Degree in
Biotechnology (Sobral). Advisor: Vicente de Paula Teixeira Pinto.
7. INCT-IBISAB Future Plans and Prospects
Based on its original design, the INCT-IBISAB surpassed several goals in this first year
of activities, especially in terms of scientific productivity and training of human resources.
Furthermore, the sum of new international networks born from scientific and technological
collaborations, such as RECODISA and MAL_ED, amplified and allowed several biomarkers
launching, including two bio-products based on transgenic milk goats and several molecular
biomarkers for the sensitive and specific diagnosis for infantile diarrhea causal agents. These
collaboration networks have also provided biomarkers for intestinal inflammation, enteropathy,
mal-absorption, permeability and intestinal damage. Regarding training on human resources we
exceeded some of the goals originally outlined for the future. The IBISAB also helped to create
the new postgraduate course between UFC and UFRJ. In this perspective, we have now, early
31
in the second year of activities, the INCT-IBISAB external call announcement for scholarships
provided by CNPq that will give significant support towards highly qualified human resources. In
the area of bioinformatics we launched web pages with interactive database for multi-centers,
thereby increasing our ability to communicate and with research crosstalk and collaborations in
local, national and international levels. The database platform developed by the INCT-IBISAB
also has inspired models for other recent international networks such as MAL_ED and
RECODISA. Also during the first year of activities, we organized and held two international
events of significant impact in science and technology, the V International Symposium XIV in
Biomedicine and Tropical Medicine Research Center and International Collaborative Infectious
Diseases Research meetings. These indicators assure us confidence that we are on the right
track to better develop and expand originally planned goals for the INCT-IBISAB. Our adopted
website agenda for multi-user laboratories allows the rational use of a modern infrastructure
and high technology imaging available to the full development of the proposed activities. Some
of the multi-user laboratories are already being successfully utilized during this first year and
others are in advanced stage, as described above. The CNPq provided resources for the INCTIBISAB rapidly and efficiently in order to hasten the management and development of our
planned activities as needed. Although barriers and challenges as to deal with bureaucratic
inertia for equipment and supplies purchase, such pitfalls did not compromise the full
achievement of goals reached in this first year, as shown in this current report. Even so and
beyond our INCT-IBISAB range, delays of more than twelve months in the legal process of
patent transfer and acquisition were seen, thus slowing-down the goals for developing new
products from them. We understand that the next activities and budget will give us far greater
support for the next following years to foster more scientific productivity, biotechnology and
training of human resources, and therefore will broaden the development of biomarkers and
bio-products toward the quality of life across the Brazilian Semiarid.
32
Appendix I – Year I Expenditures- 2009
Date
Qt. Capital
30/03/09
1 Notebook Dell vostro 1310. Dra. Gerly
31/03/09
2 No-Breaks estabilizados. Dr. Aldo
Balança analitica + Estufa microprocessado p/ cultura +
06/04/09
1 Botijão nitrogênio cap. 10,5lt. Dr. Aldo
12/05/09
2 Ar. Condicionados. Dr. Mauricio
12/05/09
2 Ar. Condicionados. Dra. Lucia
22/05/09
2 MicroComputadores. Dra. Lucia e Dra. Flavia
22/05/09
1 MicroComputador. Dr. Aldo
08/06/09
1 Capela de Exaustão de Gases pequena. Dr. Aldo
09/06/09 10 MicroComputadores. Para os laboratórios Dr. Aldo
10/06/09
1 Computador Dell. Dr. Aldo
30/06/09
1 No-Break estabilizado. Dr. Aldo
22/09/09
1 Capela de Exaustão de Gases pequena. Dra. Lucia
16/10/09
Aquisição de 2 banhos. Dra. Flavia
03/12/09
Balanças, Agitador e Phmetro. Dr. Aldo
03/12/09
Gaiolas metabólicas. Dr. Aldo
16/12/09
Cilindro de nitrogênio. Dr. Aldo
SubTotal:
Capital (Material Permanente a ser adquirido de
Date
Qt. fabricante/exportador no Exterior)
01/06/09
Central de inclusão Leica EG1160. Dra. Gerly
Microcentrifuga modelZ233 M-2, Cuba Eletroforese, Bio08/06/09
Rad Termociclador. Dra. Lucia
14/09/09
Analisador Genético - Applied. Dra. Dulciene
22/10/09
Camera Ussing - New Route. Dr. Aldo
22/12/09
Eletroencefalograma e eletroneuromiografia. Dr. Otoni
SubTotal:
Date
Qt. Costing
23/01/09
Tarifa folha de cheque
03/04/09
Manutenção de bancadas. Dr. Aldo
08/06/09
Manutenção de bancadas. Dra. Gerly
18/06/09
6 Tubos Polietileno. Dr. Armênio
13/07/09
Imposto Siscomex
13/07/09
Nitrogênio líquido (15 litros). Dra. Lucia
23/07/09
Imposto Siscomex
29/07/09
Material de consumo. Dra. Gerly
12/08/09
Toner impressora. Dr. Aldo
21/08/09
Serv. Terceiro. Manut. Elétrica Laboratório. Dr. Aldo
22/09/09
Anaerobac. Dra. Lucia
07/10/09
Transporte dos kits e microcentrifuga. Dra. Lucia
08/10/09
Glicina, Arginina e Glutamina. Dr. Aldo
16/10/09
Kits Ebiosciences. Dra. Flavia
28/10/09
Caeruleína Sulfatada. Dra. Flavia
28/10/09
Fragmento 8-37 Peptídeo. Dra. Flavia
28/10/09
Gotaq Colorless Master. Dra. Lucia
04/11/09
Corante azul de tripan. Dra. Flavia
19/11/09
Buffer´s reagentes. Dra. Lucia
19/11/09
Reagentes. Dra. Flavia
24/11/09
Impressão material bibliografico. Dr. Armênio
Year I - R$
2.794,94
7.400,00
7.961,00
3.850,00
3.850,00
3.344,01
1.672,01
2.286,18
22.940,00
3.999,94
3.850,00
2.827,00
12.800,00
6.830,00
17.700,00
1.242,00
105.347,08
Year I - R$
26.908,89
23.190,45
137.000,00
54.500,00
58.026,76
299.626,10
Year I - R$
42,00
1.490,00
1.850,00
6.820,00
40,00
240,00
80,00
7.310,40
949,11
200,00
2.716,50
300,00
249,00
9.670,00
905,00
2.037,00
712,00
262,00
2.429,26
4.140,00
133,50
Thematic
1
1
1
3
2
2
1
1
1
1
1
2
2
1
1
1
Thematic
1
2
2
1
3
Thematic
0
1
1
4
1
2
2
1
1
1
2
2
1
2
2
2
2
2
2
2
4
33
03/12/09
03/12/09
16/12/09
18/12/09
21/12/09
Eppedorf, Backer, Laminas e Luvas. Dra. Lucia
Laser Point p/ apresentação. Dr. Aldo
Reagentes (Etanol, Cloreto e Indometacina). Dra. Flavia
Buffer TAE. Dra. Lucia
Gel Pilot. Dra. Lucia
SubTotal:
1.829,00
149,00
701,00
1.563,77
445,72
47.264,26
Date
Qt. Costing
10/07/09
Frete Leica Mikrosysteme - Central Inclusão. Dra. Gerly
16/07/09
Importação Infraero - Central inclusão. Dra. Gerly
21/07/09
Frete New Route Inc - Kit´s Bio-Raid. Dra. Lucia
27/07/09
Importação Infraero - Kit´s Bio-Rad. Dra. Lucia
25/11/09
Carbono 13. Dr. Armênio
SubTotal:
Ano I - R$
1.756,62
589,55
1.777,77
490,51
22.481,10
27.095,55
Temáticas
1
1
2
2
4
Date
Qt. Costing (Pessoa Jurídica)
30/06/09
1 Serv. Terceiros. Charles Roberto S. Melo
30/06/09
1 Serv. Terceiros. Kátia Ma. Lima Nogueira
30/06/09
1 Serv. Terceiros. Antônio Haroldo P. Ferreira
24/08/09
1 Serv. Terceiros. Kátia Ma. Lima Nogueira
24/08/09
1 Serv. Terceiros. Charles Roberto S. Melo
24/08/09
1 Serv. Terceiros. Antônio Haroldo P. Ferreira
24/08/09
1 Serv. Terceiros. Francisco de Sousa Junior
30/10/09
1 Serv. Terceiros. Kátia Ma. Lima Nogueira
30/10/09
1 Serv. Terceiros. Charles Roberto S. Melo
30/10/09
1 Serv. Terceiros. Francisco de Sousa Junior
30/10/09
1 Serv. Terceiros. Antônio Haroldo P. Ferreira
30/10/09
1 Serv. Terceiros. José Amadeus Souza
30/10/09
1 Serv. Terceiros. Francisco Léo N. Aguiar
Thematic
0
0
0
0
0
0
0
0
0
0
0
0
0
SubTotal:
Year I - R$
800,00
950,00
400,00
950,00
800,00
400,00
1.000,00
950,00
800,00
500,00
400,00
400,00
752,00
9.102,00
Date
Qt. Costing
02/04/09
2 Diárias nacionais (R$187,83) Dra. Dulciene
02/04/09
1 Passagem (BH - FOR - BH). Dra. Dulciene
16/09/09
1 Passagem (FOR - RIO - FOR). Dr. Aldo
30/09/09 1 ½ Uma diária e meia Dr. Aldo
21/10/09
1 Passagem Dra. Klena (Belem x FOR x Belem)
21/10/09
1 Passagem Dr. Helder (SP x FOR x SP).
30/10/09
4 Diárias nacionais (R$187,83) Dr. Helder Tambellini
30/10/09
4 Diárias nacionais (R$187,83) Dra. Klena Silva
18/12/09
1 Passagem (FOR x BH / BH x FOR) Willy Prof Armenio
SubTotal:
Year I - R$
375,66
1.841,04
637,84
281,75
884,04
2.301,04
751,32
751,32
682,74
8.506,75
Thematic
2
2
1
1
3
3
3
3
3
Year I - R$
Thematic
Date
Qt. Costing (Fellows)
SubTotal:
Totals:
2
1
2
2
2
0,00
Year I - R$
496.941,74
34
Appendix II – Expenditures made YEAR II - 2010
Date
Qt. Capital
19/01/10
Kits para dosagem. Dra. Flavia
25/01/10
Bomba de vácuo. Dr. Aldo
22/04/10
Sala de video e tele-conferencia. IBISAB
Thematic
2
1
1
SubTotal:
Year II - R$
13.206,50
4.100,00
35.456,00
52.762,50
Thematic
4
SubTotal:
Year II - R$
72.703,49
72.703,49
Date
Qt. Costing
25/01/10
Impressão de material bibliográfico. Dr. Armênio
11/02/10
Kits de Elisa Hu IL1 e Mo TNF. Dra. Flávia
11/02/10
GoTaq Green Master Mix. Dra. Lúcia
18/02/10
Manut. Detector ED-40. Dr. Aldo
23/02/10
Manut. Nobreak´s. Dr. Aldo
23/02/10
Manut. Capela Fluxo Laminar. Dr. Aldo
16/03/10
Material de consumo. Dra. Lúcia
17/03/10
Nitrogênio liquido (10 litros). Dr. Aldo
17/03/10
Nitrogênio liquido (10 litros). Dra. Lucia
24/03/10
DNA Primer. Dr. Aldo
12/04/10
Tris Buffered e Immunblot. Dra. Flávia
SubTotal:
Year II - R$
100,00
4.835,00
1.884,00
2.100,00
515,00
180,00
4.996,60
200,00
200,00
793,65
1.500,00
17.304,25
Thematic
4
2
2
1
1
1
2
1
2
1
2
Year II - R$
Thematic
Date
Qtd Capital
22/01/10
Miografo Sistema Confocal.
Date
Qt. Costing
SubTotal:
0,00
Date
Qt. Costing
05/01/10
Frete da bomba de vácuo. Dr. Aldo
26/01/10
Frete Carbono 13. Dr. Armênio
27/01/10
Imposto Siscomex Carbono 13. Dr. Armênio
29/01/10
Serv. Terceiros. Kátia Ma. Lima Nogueira
29/01/10
Serv. Terceiros. Charles R. S. Melo
29/01/10
Serv. Terceiros. Fco. Sousa Junior
29/01/10
Serv. Terceiros. José Amadeus Souza
29/01/10
Serv. Terceiros. Thiago de Paula Silva
29/01/10
Serv. Terceiros. Ant. Haroldo P. Ferreira
01/02/10
Armazenamento Carbono 13. Dr. Armênio
01/02/10
DNA Primer´s. Dra. Lúcia
09/03/10
Impressões, encadernações Simposium. Dr. Aldo
10/03/10
Frete: Eletroencefalograma. Dr. Otoni
17/03/10
Manutenção HPLC. Dr. Aldo
24/03/10
Frete: Termocicladores. Dr. Aldo
26/03/10
Armazenamento Eletroencefalograma. Dr. Otoni
SubTotal:
Year II - R$
113,08
908,80
40,00
1.381,00
1.163,00
727,00
582,00
1.020,00
582,00
91,23
627,90
1.849,55
2.432,33
3.231,32
736,88
2.449,43
17.935,52
Thematic
1
4
4
0
0
0
0
0
0
4
2
1
3
1
1
3
Year II - R$
Thematic
Date
Qt. Costing
Passagem aerea (ida e volta) José Amadeus. FOR x
19/04/10
1 GRU x FOR. Dr. Aldo
22/04/10
2 Diárias nacionais (R$ 187,83 Cada). José Amadeus. Dr.
824,64
375,66
1
1
35
Aldo
SubTotal:
Date
Qt. Costing (Fellows)
1.200,30
Year II - R$
SubTotal:
Totals:
Thematic
0,00
Ano II - R$
161.906,06
36
Appendix III – Research Seminars IBISAB - 2009
Date
01/Jun/2009
30/Nov/200
9
19/Fev/2010
16/Dez/200
9
20/Abr/201
0
30/Abr/201
0
Lecture
“The functional
neuroimaging
and the Brain
Machine
Interface”
“Biomedical
applications of
nanostructured
materials”
“Mucosal breach
by the neonatal
immune system
in biliary atresia”
Lecturer
Prof. Draulio
Araújo
Origin
Laboratório de NeuroImagem
Funcional Departamento de Física e
Matemática Universidade de São
Paulo RP
Prof. Ricardo
Bentes de Azevedo
Coordenador do INCT de
Nanobiotecnologia da UNB
Jorge A. Bezerra,
MD
“GIS Information
System as a
research tool”
Lysozyme
Transgenic Goat
Milk:
antimicrobial
activity and
effects of
consumption
“Molecular
epidemiology of
gastrointestinal
virus”
John Nuckols, PhD
William and Rebecca Balistreri Chair
of Pediatric Hepatology, Professor of
Pediatrics, Director, Digestive Health
Center Director of Research, Division
of Gastroenterology, Hepatology and
Nutrition Cincinnati Children's
Hospital Medical Center
State Colorado University and
FIC/NIH
Elizabeth A. Maga,
PhD
Associate Research Biologist
Department of Animal Science
University of California, Davis
José Paulo
Gagliardi Leite,
PhD
Laboratório de Virologia Comparada
e Ambiental Fundação Oswaldo Cruz
- Ministério da Saúde
37
Biomarkers and Bioproducts Developed and / or Validated
Tables 1-8 below describe the developed and / or validated bio-markers designed by INCT-IBSAB and available for use as DNA
probes for the diagnosis of intestinal infection, determination of virulence factors and biological studies in the Institute and intrinsic
goals in two international networks, RECODISA and MAL_ED, as described in previous sections.
Table 1: Bio-markers developed for the diagnosis and study of bacterial virulence genes.
Biomarcadores
Target gene description
Gene for diagnosis
Primers sequence (5´- 3´)
Product (pb)
Amplificação
S: ATTGTCCTCAGGCATTTCAC
AS: ACGACACCCCTGATAAACAA
215
20’’ a 95ºC, 20’’ a 57ºC, 1’ a 72ºC
S: ATGAAATTAAAACAAAATATCGA
AS: TCATTGGCTTTTAAAATAAGTCAA
798
30’’ a 95ºC, 30’’ a 58ºC, 1’ a 72ºC
S: ATGATGGCTTCTTCGGATAG
AS: GCTCCTATGCTTACAACTGC
176
S: GGTATGATTTCTACAAGCGAG
AS: ATAAAAGACTATCGTCGCGTG
502
S: TAGAGAGATAGCAAAAGAGA
AS: TACACATAATAATCCCACCC
251
S: TGACAGTGGATCAGGCGTGT
AS: TTCTGTGCGCCAAGAATGAC
558
S: TTCAGCCGAAAGACGAAATCG
AS: TCTGCGCATTCATACCAACAT
518
astA b,d – toxina termo-estável agregativa, EAST1,
8 EAEC
S: GCCATCAACACAGTATATCCGA
112
9 aap b,d – proteína anti-agregativa, dispersina, EAEC
AS: CGATATTATTTAACCCATTCGG
356
aaiC a,c – ilha ativadora do gene aggR, E. coli
1 enteroagregativa (EAEC)
b,c
2 aggR – regulador de aderência agregativa, EAEC
a,c
3 hipO – hipurato hidrolase de C. jejuni
a,c
4 ask – aspartato quinase de C. coli
5 glyA a,c – serina hidroximetiltransferase de C. lari
Virulence gene
b,c
6 pet – toxina codificada por plasmídio, EAEC
a,d – proteína envolvida em colonização, EAEC
7 pic
30’’ a 95ºC, 30’’ a 51ºC, 45" a 72ºC
30’’ a 95ºC, 30’’ a 46ºC, 45" a 72ºC
30’’ a 95ºC, 30’’ a 55ºC, 1’ a 72ºC
30’’ a 95ºC, 30’’ a 55ºC, 1’ a 72ºC
Continuation of Table 1: Bio-markers developed for the diagnosis and study of bacterial virulence genes.
Target gene description
Biomarkers
10 cdtA a,d – toxina citoletal distensora (CDT) porção, C.
jejuni
Primers sequence (5´- 3´)
Product (pb)
S: TTGGCGATGCTAGAGTTTGG
AS: ACCGCTGTATTGCTCATAGGG
175
S: CTCGCGTTGATGTAGGAGCTA
AS: GCAGCTAAAAGCGGTGGAGTA
418
S: AGCCTTTGCAACTCCTACTGG
AS: GCTCCAAAGGTTCCATCTTC
270
S: AGCTGCTTCGCAACTTTCTACGGT
AS: TGCACTCTCGGCTGCAAAGTCT
325
S: AAGAGTGAGGCGAAATTCCA
AS: GCAAGATGGCAGGATTATCA
385
S: TCATGCGGTGGCATTAGAATGGG
AS: GCGACCGATGATAACATCAAGGCT
658
S: TGGGGCTTCAAATCGGTGCTGA
AS: GCGACCGATGATAACATCAAGGCT
326
S: AGGCTTTGGCTCATCACGTCG
AS: GGTCGCTTCACCGCGTATGG
574
S: TCAGAAGCTCAGCGAATCATTG
AS: CCATTATCACCAGTAAAACGCACC
931
11 cdtB a,d – CDT porção B, C. jejuni
12 cdtC a,d – CDT porção C, C. jejuni
13 flaA a,d – flagelina A, C. jejuni
14 pldA a,d – fosfolipase A, C. jejuni
15 ciaB a,d – antígeno de invasão de Campylobacter
16 racR a,d – regulador de resposta DNA-ligante, C. jejuni
17 dnaJ a,d – proteína chaperona DnaJ, C. jejuni
18 SAT a,c – autotransportador de toxina secretada,
patotipos de E. coli
Amplificação
30’’ a 95ºC, 30’’ a 56ºC, 45" a 72ºC
30’’ a 95ºC, 30’’ a 59ºC, 45" a 72ºC
30’’ a 95ºC, 30’’ a 64ºC, 45" a 72ºC
30’’ a 95ºC, 30’’ a 58ºC, 1' a 72ºC
Legend:
a Gene cromossomal; b Gene plasmidial, c PCR comum; d PCR Multiplex.
S: iniciador senso; AS: iniciador anti-senso
pb: pares de bases
39
Table 2: Validated bio-markers from the INCT_Biomedicine and their use in international networks RECODISA and MAL_ED.
Biomarkers
Target gene description
Gene for diagnosis
1 ipaH
2
a,c
– E. coli enteroinvasiva (EIEC)
aatA b,c – proteína transportadora antiagregação, E. coli enteroagregativa (EAEC)
stx1 a,d – shiga-toxina 1, E. coli
3 enterohemorrágica (EHEC)
4 stx2
a,d –
shiga-toxina 2, EHEC
eae a,d – intimina, E. coli enteropatogênica
5 (EPEC)
6 bfpA
b,d –
proteína bfpA, EPEC
LT b,d – toxina termolábil, E. coli
7 enterotoxigênica (ETEC)
8 ST
b,d –
toxina termoestável, ETEC
Primers sequence (5´- 3´)
Product (pb)
S: TGGAAAAACTCAGTGCCTCT
AS: CCAGTCCGTAAATTCATTCT
423
S: CTGGCGAAAGACTGTATCAT
AS: CAATGTATAGAAATCCGCTGTT
630
S: CAGTTAATGTGGTGGCGAAGG
AS: CACCAGACAATGTAACCGCTG
348
S: ATCCTATTCCCGGGAGTTTACG
AS: ATCCTATTCCCGGGAGTTTACG
584
S: CCCGAATTCGGCACAAGCATAAGC
AS: CCCGGATCCGTCTCGCCAGTATTCG
881
S: GGAAGTCAAATTCATGGGGGTAT
AS: GGAATCAGACGCAGACTGGTAGT
300
S: CACACGGAGCTCCTCAGTC
AS: CCCCCAGCCTAGCTTAGTTT
508
S: GCTAAACCAGTAGAGGTCTTCAAAA
AS: CCCGGTACAGAGCAGGATTACAACA
147
S: ATGAAAAAAATTAAGTTTGTTATCTT
356
AS: GGTCGCGAGTGACGGCTTTGT
112
PCR conditions
Reference
30’’ a 95ºC, 30’’ a
57ºC, 45" a 72ºC
MalEd Project,
2009.
30’’ a 95ºC, 30’’ a
55ºC, 1’ a 72ºC
Sheik et al, 2002.
Virulence gene
9 aap b,d – dispersina, EAEC
10 astA
b,d
– EAST1, EAEC
Piva et al, 2003.
Legend:
a Gene cromossomal; b Gene plasmidial, c PCR comum; d PCR Multiplex.
S: iniciador senso; AS: iniciador anti-senso
pb: pares de bases
Referrences:

Sheikh J, Czeczulin JR, Harrington S, et al. A novel dispersin protein in enteroaggregative Escherichia coli. J Clin Invest 2002; 110:1329-37.

Piva IC, Pereira AL, Ferraz LR, et al. Virulence markers of enteroaggregative Escherichia coli isolated from children and adults with diarrhea in Brasilia, Brazil. J Clin
Microbiol 2003; 41:1827-32.
40
Table 3: Bio-markers developed in INCT_Biomedicina for diagnosis and assessment of genetic polymorphism.
Target gene description
Gene for diagnosis
Biomarkers
Primers sequence (5´- 3´)
1 Análise de polimorfismo de um único nucleotídeo no
estudo de Intolerância à Lactose (C/T -13910)
2 Análise de polimorfismo de um único nucleotídeo para o
gene do Receptor Toll-Like-5
S: GAGTGTAGTTGTTAGACGGAGAC
AS: ATCAAACATTATACAAATGCAAC
S: GGTAGCCTACATTGATTTGC
AS: GAGAATCTGGAGATGAGGTACCCG
Product (pb)
Amplification
210
30’’ a 95ºC, 30’’ a 54ºC, 1’ a 72ºC
277
30’’ a 95ºC, 30’’ a 60ºC, 1’ a 72ºC
Table 4: Bio-markers validated for the study of polymorphisms in regulatory genes of cytokines.
Biomarkers
Target gene description
Gene for diagnosis
1 Análise de polimorfismo de um único
nucleotídeo para o gene de Interleucina
8 humana (IL-8)
2 Análise de polimorfismo de um único
nucleotídeo no estudo de Intolerância à
Lactose (G/A -22810)
Amplification
Reference
Primers sequence (5´- 3´)
Product (pb)
S: ACTATATCTGTCACATGGTACTATG
AS: CTTATCAAATACGGAGTATGACG
166
30’’ a 95ºC, 30’’ a 54ºC, 1’ a 72ºC
Jiang et al.,
2003
S: AACAGGCACGTGGAGGAGTT
AS: CCCACCTCAGCCTCTTGAGT
448
30’’ a 95ºC, 30’’ a 60ºC, 1’ a 72ºC
Bünning et
al., 2003
References:

JIANG, Z. D. et al. Genetic susceptibility to enteroaggregative Escherichia coli diarrhea: polymorphism in the interleukin-8 promotor region. J. Infect. Dis. 2003; 188:
506-511.

BÜNING, C et al. The C/C (-13910) and G/G (-22018) genotypes for adult-type hypolactasia are not associated with inflammatory bowel disease. Scand J Gastroenterol
2003; 38:538-542.
Table 6: Bio-markers validated for the study of intestinal glutamine transport.
Biomarkers
Target gene description
Sequência dos iniciadores (5´- 3´)
Product (pb)
Amplification
Reference
Gene for diagnosis
1
2
Transportador de aminoácidos
intestinal (SN-1) para coelho
Transportador de aminoácidos
intestinal (SN-2) para coelho
S: GGCAGGGGTTTCCTACAGA
AS: CGATGTCTTCCCCTCGAA
S: CTGGGACAGAGGGCATTC
AS: CGGATTTGATGATGAACAGGT
69
106
20’’ a 95ºC, 20’’ a 62,2ºC, 45’’ a 72ºC
20’’ a 95ºC, 20’’ a 62,2ºC, 45’’ a 72ºC
Talukder et al.,
2008
References:

Talukder et al. Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter. Am J Physiol Gastrointest Liver Physiol
2008; 294: G1301–G1310.
41
Table 5: Bio-markers developed to study transport proteins and joints strong in intestinal barrier function.
Biomarkers
Target gene description
Primers sequence (5´- 3´)
Product (pb)
Amplification
S: TCTACGAGGGACTGTGGATG
AS: TCAGATTCAGCAAGGAGTCG
84
20’’ a 95ºC, 20’’ a 55ºC, 45’’ a 72ºC
S: CCCACCACCACCAGCTTAAT
AS: GAAATGGCTTCCAGGTCAGC
170
20’’ a 95ºC, 20’’ a 60ºC, 45’’ a 72ºC
S: AGGACTTCCTGCTGACATCCA
AS: TCCACCCACTACAGCCACTCT
154
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
S: GACCATCGCCTACGGTTTGA
AS: AGGTCTCGGGGATGCTGATT
116
20’’ a 95ºC, 20’’ a 60ºC, 45’’ a 72ºC
S: CTCGCACGTATCACAAGCTGA
AS: CCTCAGGATATGGCTCCTTCC
137
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
S: AAGAGCAGCCAAAGGCTTCC
AS: CGTCGGGTTCACTCCCATTA
199
20’’ a 95ºC, 20’’ a 60ºC, 45’’ a 72ºC
S: AACAGCCCCCTAATGTGGAAG
AS: GAGTAGGCCATTGGACTGTCG
112
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
S: TGGTGTCTGCCATTGTACGC
AS: CCACTGGTGACCCAAGCATT
165
20’’ a 95ºC, 20’’ a 60ºC, 45’’ a 72ºC
S: CTGGCAGCAGCAATCTTACCT
AS: AAAGGACTTGGGAGGTGTCCA
119
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
S: CCTGAAGAAGATGACCGTTGG
AS: GCTGGGGAAGACTGGAAGAGT
103
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
S: AGCCTCGTCCCGTAGACAAA
AS: TGAATTTGCCGTGAGTGGAG
183
20’’ a 95ºC, 20’’ a 60ºC, 45’’ a 72ºC
S: GTTACCAGGGCTGCCTTCTCT
AS: AACTTGCCGTGGGTAGAGTCA
116
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
13 Transportador de peptídeo intestinal-1 (PEPT-1)
para coelho
S: GGGAGTCTGCTGTCCACAATC
AS: GTACATCCCACTGCCGATGAT
153
20’’ a 95ºC, 20’’ a 64,6ºC, 45’’ a 72ºC
14 Co-transportador Sódio-Glicose intestinal para
coelho (SGLT-1)
S: CTGACTGGGTTCGCTTTTCAC
AS: GCATCTCGGAAGATGTGGAAG
155
20’’ a 95ºC, 20’’ a 65ºC, 45’’ a 72ºC
S: GCCGTGGGCAAGGTCATCCC
AS: GCAGCTTTCTCCAGGCGGCA
113
20’’ a 95ºC, 20’’ a 64,6ºC, 45’’ a 72ºC
1 Claudina-1
2 Claudina-2 para camundongo
3 Claudina-2 para rato
4 Zona Ocludens-1 para camundongo
5 Zona Ocludens-1 para rato
6 Ocludina para camundongo
7 Ocludina para rato
8 Beta-catenina para camundongo
9 Beta-catenina para rato
10 Transportador de peptídeo intestinal-1 (PEPT-1)
para rato
11 GAPDH para camundongo
12 GAPDH para rato
15 GAPDH para coelho
42
Table 7: Bio-markers developed for the study of receptor guanylyl cyclases and natriuretic peptide C.
Target gene description
Biomarkers
1 Receptor de Guanilato ciclase A de rato (GC-A)
2 Receptor de Guanilato ciclase B de rato (GC-B)
3 Receptor de Guanilato ciclase C de rato (GC-C)
4 Receptor de Peptídeo Natriurético C de rato (NPR-3)
5 RNA ribossômico cadeia 18S de rato (18S rRNA)
Primers sequence (5´- 3´)
Product (pb)
Amplification
S:GAACCGAAGCTTCCAAGGTG
AS: GTGGATATCCCAGAGGCCAGT
218
30’’ a 95ºC, 30’’ a 58ºC, 1' a 72ºC
S: CATCTGCATCGTCACCGAGT
AS: TCCACCACGCAGTTAGAGGAC
186
30’’ a 95ºC, 30’’ a 59ºC, 1' a 72ºC
S: GGCGGGATACAATCCAGAGAG
AS: ACGGTGCCGTAGAACTTGGTC
166
30’’ a 95ºC, 30’’ a 63ºC, 1' a 72ºC
S: CCTACAATTTCGACGAGACCAAA
AS: TCGCTCACTGCCCTGGAT
201
30’’ a 95ºC, 30’’ a 59ºC, 1' a 72ºC
S: ACATCCAAGGAAGGCAGCAG
AS: GCTGGAATTACCGCGGCTG
179
30’’ a 95ºC, 30’’ a 60ºC, 1' a 72ºC
Table 8: Bio-markers developed for the diagnosis of bacterial resistance.
Target gene description
Primers sequence (5´- 3´)
Biomarkers
Gene for diagnosis
1 katG - catalase-peroxidase, Mycobacterium
tuberculosis
S: GTCGGCGGTCACACTTTC
AS: GCTACCACGGAACGACGAC
2 rpoB – subunidade beta da RNA polimerase
direcionada ao DNA, M. tuberculosis
S: TACGGTCGGCGAGCTGATCC
AS: TACGGCGTTTCGATGAACC
3 inhA - enoil-(acil proteína carreadora)
redutase, M. tuberculosis
S: TGCTCGAACTCGACGTGCAA
AS: CGAAGCATACGAATACGCCGA
Product (pb)
PCR conditions
86
30’’ a 95ºC, 30’’ a 64ºC, 1’ a 72ºC
165
30’’ a 95ºC, 30’’ a 61ºC, 1’ a 72ºC
209
30’’ a 95ºC, 30’’ a 62ºC, 1’ a 72ºC
43
44
45
46