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Simpósio Comemorativo dos 50 anos do
Programa de Pós-Graduação em Ciências
(Bioquímica) da UFPR
LIVRO DE RESUMOS
2015
ii
Universidade Federal do Paraná
Programa de Pós-Graduação em Ciências (Bioquímica)
Simpósio Comemorativo dos 50 anos do Programa de Pós-
Graduação em Ciências (Bioquímica) da UFPR
Livro de Resumos
Curitiba-PR
2015
iii
Programa de Pós-Graduação em Ciências (Bioquímica)
REALIZAÇÃO:
Coordenação do Curso dePós-Graduação em Ciências (Bioquímica):
Glaucia Regina Martinez
Emanuel Maltempi de Souza
Chefia de Departamento:
Rose Adele Monteiro
Joana Léa Meira Silveira
Colegiado de Curso de Pós-Graduação:
David Alexander Mitchell
Sheila Maria Brochado Winnischofer
Miguel Daniel Noseda
Guilherme Lanzi Sassaki
Marcelo Muller Dos Santos
Leonardo Magalhães Cruz
Maria Eliane Merlim Rocha
Nadia Krieger
Alessandra Biz (Discente)
Alexsandro Vinícius Nogueira (Discente)
Rocio del Pilar Cuaspa Ropain (Discente)
Apoio discente na organização das atividades do evento:
Paloma Bonato
Sarah Sacks Timoteo
Maura Harumi Sugai
Ester Mazepa
Rafaela Perez
Shayla Fernanda Barbieri
Heloisa Bruna Soligo Sanchuki
Carlos Eduardo Sanchuki
Edileusa Cristina Marques Gerhardt
Vanessa Kessler Chicora
Manuel Jose Pinero Gavida
Fernanda Gravina
Montagem e organização do livro de resumos:
Otávio Martins Cruz (Discente)
Patrícia da Silva Peres (Discente)
iv
Programa de Pós-Graduação em Ciências – Bioquímica
APRESENTAÇÃO
Simpósio Comemorativo dos 50 anos do Programa de
Pós-Graduação em Ciências (Bioquímica) UFPR
O Simpósio Comemorativo dos 50 Anos visa fortalecer a interação dos
docentes e pós-graduandos, motivar os jovens pesquisadores, promover maior
integração entre pesquisadores de diferentes Instituições Nacionais e
Internacionais e enriquecer a formação dos pós-graduandos e docentes.
O Programa de Pós-Graduação em Ciências (Bioquímica) - PPGBq (conceito
6, triênio 2010-2012) agrega linhas de pesquisa bem estabelecidas e
diversificadas na área de bioquímica básica e aplicada com importante
inserção estadual e nacional.
APOIO:
PATROCÍNIO:
Programa de Pós-Graduação em Ciências (Bioquímica)
5 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
SUMÁRIO
APRESENTAÇÃO ................................................................................................................. iv
SUMÁRIO ............................................................................................................................... 5
RESUMOS .............................................................................................................................. 6
ÍNDICE DE AUTORES ........................................................................................................ 61
ANEXO .................................................................................................................................. 65
PROGRAMAÇÃO DO EVENTO ..................................................................................... 65
Programa de Pós-Graduação em Ciências (Bioquímica)
6 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
RESUMOS
ANTI-INFLAMMATORY EFFECTS OF THE POLYSACCHARIDES PRESENT
IN THE BLACKBERRY WINE
Caillot, A. R. C1; Bezerra, I. L.; Santana-Filho, A. P.; Sassaki, G. L.
Biochemistry and Molecular Biology Department, Federal University of Parana,
Curitiba, Brazil
Introduction: Blackberry wine is recognized as a natural source of essential
minerals and many bioactive phytochemicals that can play an important role in
health promotion. This work aims to evaluate receptor mediated anti-
inflammatory activity of the polysaccharides from blackberry wine. Methods:
The blackberry wine were precipitated by addition of EtOH-(3V) and
centrifuged. The precipitate was then dialyzed generating polysaccharide
fraction (PVA). Those were submitted to freeze-thawing and centrifugation,
resulting in soluble (PVAS) and insoluble (PVA-I) fractions. The fraction PVAS
was submitted the Fehling treatment, resulting in two new fractions: supernatant
(PVAFESB) and precipitate (PVAFEPPT). The fractions were hydrolyzed with
TFA at 100°C/14h. Thereafter the sample was dried and reduced with NaBH4
giving rise to alditols, which were acetylated. The resulting alditol-acetates were
analyzed by GC–MS. The fraction PVAS was evaluated regarding anti-
inflammatory activity. RAW-264.7 cells were treated with LPS alone (1µg/ml) or
in combination with PVAS (100µl/ml) for 24h. After 24h, anti-inflammatory
activity was evaluated on culture supernatants using Quanti-Blue and following
manufacture´s recommendations. Results: The monosaccharide composition of
the PVAS showed Man-(33.1%), Glc-(12.6%), Gal-(19.1%), Ara-(12.9%), Rha-
(5.5%), Xyl-(2.8%), Fuc-(1.3%) and GalpA-(12.9%). 1H/13C HSQC NMR
analysis of the fraction PVAFESB showed signals at chemical shift (C1/H1)
103.2/4.47, which are characteristic of (1→3)-linked b-D-Galp units present in
arabinogalactan-II. The fraction PVAFEPPT 1H/13C HSQC showed correlations
corresponding to a (1→6)-linked a-D-mannan, with chemical shift at 99.08/5.09
and 98.6/4.95. We also demonstrated that fraction PVAS inhibited receptor
mediated LPS triggered inflammation on RAW-264.7 cells using a reporter gene
assay based on NF-kb transcriptional activity.
Financial Support: CAPES
Programa de Pós-Graduação em Ciências (Bioquímica)
7 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
STRUCTURAL CHARACTERIZATION OF POLYSACCHARIDES
EXTRACTED BY INFUSION OF SEDUM DENDROIDEUM LEAVES
Oliveira A.F., Cordeiro L.M.C., Iacomini, M. and Cipriani, T.R.
Biochemistry and Molecular Biology Department, Federal University of Parana,
Curitiba, Brazil
Sedum dendroideum is a medicinal plant known in Brazil as bálsamo. Its leaves
are traditionally used to treat skin inflammations and gastric disorders. Infusion
of its lyophilized leaves (1.30 kg) was used to extract polysaccharides, which
were fractionated by freezing/thawing process, dialysis and anion exchange
chromatography. The cold water soluble fraction (6.66 g) was dialyzed with a
100 kDa cut-off membrane, yielding the fractions Rsbal (2.55 g), Esbal-I (3.35
g) and Esbal-II (0.55 g). Rsbal and Esbal-II showed homogeneous HPSEC
profiles. Monosaccharide analysis showed that Esbal-II contained GalA
(82.3%), Ara (4.5%) and Gal (4.2%), suggesting the presence of a
homogalacturonan, whereas Rsbal contained GalA (47.2%), Ara (23.2%), Gal
(24.8%) and Glc (4,9%), suggesting the presence of a homogalacturonan and
an arabinogalactan. Rsbal (100 mg) was subjected to anion exchange
chromatography yielding the fractions Rsbal-H2O (25 mg), Rsbal 0,5M (62.1
mg) and Rsbal-1M (1.2 mg). Rsbal-H2O showed GalA (60%), Gal (25.3%), Rha
(1.9%), Ara (3.9%) and Glu (6.8%). This acid polysaccharide was eluted with
H2O, probably because its GalA units are methyl-esterified. Rsbal-0,5M
presented GalA (62.1%), Gal (17.4%), Ara (17.1%) and Rha (1.0%), suggesting
the presence of a homogalacturonan and an arabinogalactan. Its NMR analysis
(HSQC) showed chemical shifts at 103.3/4.50 of C1/H1 of β-D-Galp;
106.5/5.80, 107.9/5.42 and 108.9/5.25 of C1/H1 of α-L-Araf; 100.2/4.82 and
99.0/5.15 of C1/H1 of 6-OMe-α-D-GalpA and α-D-GalpA, respectively;
52.9/3.81 of -COO-CH3; and 70.5/5.11 and 71.3/4.78 of C5/H5 of 6-OMe-α-D-
GalpA and α-D-GalpA, respectively. Methylation analysis will be performed to
obtain more structural informations.
Programa de Pós-Graduação em Ciências (Bioquímica)
8 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
MMP14 GENE REGULATION BY DNA METHYLATION IN THE INTRONIC
REGION
Chequin, A.1; Manica, G. C. M.1; Klassen, L. M. B.1; Ramos, E. A. S.1; Toledo,
M. B.1; Brandão, Y. O.; Souza, E. M.2; Klassen, G.1
1Dep. de Patologia Básica, Laboratório de Epigenética. Universidade Federal
do Paraná. UFPR, PR, Brasil.
2: Dep. de Bioquimica. Universidade Federal do Paraná. UFPR, PR, Brasil.
Metastases are responsible for 90% of deaths in breast cancer. The
metalloproteinase 14 (MMP-14) is an important protein related to metastatic
process, and present three CpG islands (CGIs), one on the promoter region,
which known regulates gene expression, and two located in intronic regions,
that haven´t been studied. Recent reports showed that methylation on intronic
regions may be important to regulate the human transcriptome. The aim of this
study was evaluate the effect of DNA methylation inhibitior (DAC) and inhibitor
of histone deacetylase (TSA) in the demethylation of intronic islands of MMP14
in breast tumor cells lines. The expression of the MMP14 in two breast tumor
cell lines was evaluated by qRT-PCR. The regions containing the CGIs were,
after sodium bisulfite treatment, cloned and sequenced. The cell line MMP14
negative was treated with DAC, TSA, or both, and also subjected to qRT-PCR
and sequencing. MCF7, which shows no expression of MMP14, showed high
levels of methylation in CGIs, as well as PMC-42, which expresses MMP14,
showed lower levels of methylation. After treatments, MCF7 passed to express:
2.6 (DAC), 3.2 (TSA) and 2.9 (DAC+TSA) more MMP14 than mock in its three
CGIs respectively, and the rates of methylation in first CGI had a reduction of
40,7%, 18.7%, 66%. On the intronics islands 2 and 3, these reductions were
more pronounced: 95%, 59%, 100% and; 81.3%, 64%, 73.3%, in the same
treatments. Apparently, MMP14 is being regulated by methylation on CGIs
located in first íntron in addition to promoter region.
Programa de Pós-Graduação em Ciências (Bioquímica)
9 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
SYD-1 INDUCE RESPIRATION DYSFUNCTION ON HEPG2 WITH
OXIDATIVE PHOSPHORYLATION PATHWAY ACTIVATED
Brandt, A.P.1; Pires, A. R. A. 1; Echevarria, A.2; Canuto, V.C.2; Cadena,
S.M.S.C.1
1Departamento de Bioquímica e Biologia Molecular, UFPR, PR;
2Departamento de Química,UFRRJ, RJ. Brazil.
An important antitumor effect of SYD-1 (3-[4-chloro-3-nitrophenyl]-1,2,3-
oxadiazolium-5-olate) has been shown. We previously reported that SYD-1
impairs oxidative phosphorylation in isolated mitochondria. The expressive
inhibition of electron transport throughout the respiratory chain was related with
its antitumor effect. Now, to better visualize the effects of SYD-1 under
metabolic parameters, with highlight to the mitochondrial function, we evaluated
the effects of this derivative on human liver carcinoma cells (HepG2) cultured
with free glucose DMEM medium supplemented with galactose and glutamine
(aiming to induce these cells to obtain energy through oxidative respiration).
SYD-
after 24h of treatment, as evaluated by MTT assay. Using crystal violet staining,
after 72h of treatment, SYD-1 decreased the viability of these cells by ~35% to
as MTT assay after 24h of treatment, reaching only ~43% to the high
inhibited by SYD-1. However, the compound strongly inhibited the leak state to
all concentrations used (15-
cultured with DMEM medium supplemented with high concentration of glucose
(previous data), these results were less pronounced indicating SYD-1 effects
may be related to impairment of oxidative respiration.
Financial Support: CNPq, CAPES, INCT-Redoxoma
Programa de Pós-Graduação em Ciências (Bioquímica)
10 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ACTIVITY OF HEXAHYDROXYTRIPHENYLENE ON CELL VIABILITY AND
REACTIVE OXYGEN SPECIES LEVELS IN HUMAN GLIOMA T98G CELLS
Ribeiro, C.S.P¹; Cruz, O.M¹; Pino-Gomes, R.¹; Bark, J.M¹; Winnischofer, H.²;
Martinez, G.R¹; Winnischofer, S.M.B¹
¹ Departamento de Bioquímica e Biologia Molecular – Universidade Federal do
Paraná, Paraná, Brasil.
² Departamento de Química – Universidade Federal do Paraná, Paraná, Brasil.
Glioblastoma multiform is considered the most aggressive cancer of the central
nervous system, with a very high mortality rates. This scenario shows the
inefficiency of the current treatment, which is based on the use of temozolomide
chemotherapy followed by radiation therapy. Therefore, the aim of this study is
to evaluate the effects of hexahydroxytriphenylene (HHTP) on cell viability and
intracellular reactive oxygen species (ROS) levels in T98G cell line Cell viability
was assessed by MTT method, testing different concentrations of HHTP (10,
25and 50 μM) and drug exposure times (12, 24 and 48 hours). After 12 hours, it
was observed that the HHTP treatment did not alter cell viability at all times and
concentrations tested. After 24 hours exposure of 50μM HHTP, the viability of
the cells was reduced by 25%. At the time of 48h the reduction in the viability
was 44% and 46% with 25 and 50 μM HHTP, respectively. In addition, it was
also evaluated the levels of ROS by the oxidation of the fluorescent probe 2 ', 7'
- diclorodiidrofluorescina (DCFH-DA). Results showed that T98G cells treated
with HHTP at 50 μM for 12 hours displayed an increase in the ROS levels, by
40%. After 24 hours, it was observed a significant increase in ROS levels, at all
concentrations HTTP. In 48 hours, persistent high ROS levels were observed,
reaching 30% at 25 μM HHTP and 47% at 50 μM HHTP. Together, these data
suggest that treatment with HHTP in glioblastoma model, reduces the cell
viability by concentration-dependent, accompanied by increase in ROS levels.
Financial Support: CNPq, CAPES, INCT-Redoxoma
Programa de Pós-Graduação em Ciências (Bioquímica)
11 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
DETERMINING THE BACTERIAL DIVERSITY IN THE MANGROVE SOIL IN
THE REGION OF PARANAGUÁ BAY, BRAZIL, AND THE NATURAL
FACTORS AFFECTING IT
Denny Marcel Seccon, Daniel Renato Lammel, Eduardo Balsanelli, Emanuel
Maltempi de Souza, Helisson Faoro, Michelle Zibetti Tadra Sfeir, Paulo da
Cunha Lana, Roseli Wassem, Fabio de Oliveira Pedrosa
Departamento de Bioquímica e Biologia Molecular – Universidade Federal do
Paraná, Paraná, Brasil.
Mangroves are unique habitats present in the interface of sea and land. The
peculiar characteristics of this environment are likely due to fluctuation in sea
salinity and to the anaerobicity of the soil. Mangroves provide shelter for many
terrestrial and sea animals, protect the shores from erosion and are well suited
for economic exploitation. As expected, the microorganisms inhabiting its
sediments play a pivotal role in maintaining its functionality and health by
interacting with the plants growing thereby. A better understanding of the
microbial communities of mangroves has become increasingly important since
mangroves have been strongly affected by human activities and pollution. This
work characterizes the biodiversity of the prokaryotic community living in the
mangrove soil of the region of Paranaguá Bay, Paraná, Brazil, depicting its
general profile and which environmental factors contribute to its constitution,
such as seasonal changes, the proximity to open sea, the rhizosphere of the
prevalent plants and the physicochemical parameters of the soil.
Financial Support: CAPES
Programa de Pós-Graduação em Ciências (Bioquímica)
12 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ANTI-METASTATIC AND ANTIINFLAMATORY ACTIVITY OF A L-
GALACTAN TYPE POLYSACCHARIDE ISOLATED FROM A ASCIDIA
STYELA PLICATA
Diana C. Restrepo E.1, Felipe C.O.B. Teixeira2, Eliene O. Kozlowski 2, Jhonny
Colorado Ríos1, Alejandro Martinez1, Mauro S.G. Pavão2
1Universidad de Antioquia, Medellín, Colombia; Research group of Marine
Natural Products, Faculty of Pharmaceutical Chemistry.
2Universidade Federal do Rio de Janeiro, Brazil; Research group of
Biochemistry and Cellular Biology of Glycoconjugates, Biochemistry Institute.
Ascidians (Tunicata) are sessile marine invertebrates. Previous studies have
accessed novel biological activities for a sulfated polysaccharide isolated from
the tunic of the Brazilian ascidian Styela plicata. This carbohydrate is a high
molecular weight 3- -galactopyranosyl. The ability of a
cancer cell to metastasize successfully depends on its individual properties and
the properties of immune, bloodstream and lymphatic cells. It is known that
Heparin treatment attenuates metastasis and inflammation through inhibition of
P-selectin, present on platelets and neutrophil, binding to its ligands on tumor
cells. In this preliminary study, in vitro and in vivo assays were performed to
evaluate the polysaccharide cytotoxicity on LLC cell line, and its anti-
inflammatory and antimetastatic effect with the purpose of finding new therapies
to treat these diseases.
Financial Support: COLCIENCIAS (Administartive Department of Science,
Technology- Colombia)
Programa de Pós-Graduação em Ciências (Bioquímica)
13 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
Β-(1→4)-GALACTAN FROM SICANA ODORIFERA MODULATING TO
MACROPHAGES FOR IMMUNOSSUPRESSOR PROFILE
Abreu, E. C. A.; Noleto, G. R.
Departamento de Bioquímica e Biologia Molecular – Universidade Federal do
Paraná, Paraná, Brasil.
Polysaccharides from different plants have great potential of application. The
effects of β-(1→4)-galactan from fruit of S. odorifera on mice peritoneal
macrophages were investigated. The results show that in 24h of incubation
100µg/ml of the polymer reduced the cell viability of macrophages at 20%. In
contraste, an increase of 25% and ~32% with 50 µg/ml and 100 µg/ml,
respectively in 48h was observed. The treated groups with 50 µg/ml and 100
µg/ml of the β-(1→4)-galactan decreased at ~47% and 67%, respectively, the
macrophages number with activation morphologic profile. The phagocytic
activity was reduced in ~50% with 50µg/ml. When the macrophages were
incubated with LPS (100ng/ml) + β-(1→4)-galactan the phagocytosis was
reduced at ~40% and ~33% (50 µg/ml and 100 µg/ml, respectively) in
comparison with the cells treated with only LPS. In the presence of LPS
(100ng/ml) + polymer (50 and 100 µg/ml) during 1h, the superoxide anion
production by macrophages was reduced at 70%). The production of nitric
oxide in 48h by these cells in presence of LPS 100 ng/ml + 50 µg/ml of β-
(1→4)-galactan was reduced in ~20%. The pro- and anti-inflammatory
interleukins production was also evaluated. In 6h of incubation, macrophages
treated with β-(1→4)-galactan (50 µg/ml) increased at ~100% the TNF-α level.
But 100µg/ml of the polymer in 48h reduced at ~95% the IL-1β production. In
the same time of incubation, the β-galactan polymer (100µg/ml) increased at
~57% the IL-10 level. Taken together, the results of this study show that the β-
(1→4)-galactan of S. odorifera trigger an immunossupressor profile.
Financial Support: CNPq, CAPES, INCT-Redoxoma
Programa de Pós-Graduação em Ciências (Bioquímica)
14 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
CHEMICAL MODIFICATION OF AGAROSE FOR THE DEVELOPMENT OF
NEW BIOMATERIALS
Estela M. Aranha, Miguel D. Noseda, Janaina G. Heuke, Diogo R. B. Ducatti,
Alan G. Gonçalves, Maria Eugênia D. Noseda
Laboratório de Glicobiologia Estrutural de Carboidratos - Algas Marinhas
(GLICAM).
Departamento de Bioquímica e Biologia Molecular, Setor de Ciências
Biológicas, Universidade Federal do Paraná, Paraná, Brasil.
The search for new materials from renewable and biodegradable sources has
increased, and red algae polysaccharides, for example agarose, have drawn
attention, due to its wide application and abundance. Inserting good leaving
groups in polysaccharides by semi-synthesis can transform them into potential
precursors in the development of new materials with technological and
pharmaceutical applications, such as biotechnology field. The objective of this
study was the inclusion of tosyl groups in agarose. The synthesis of tosyl
agarose was started by reducing the polysaccharide terminals with NABH4,
followed by tosylation in heterogeneous aqueous medium (yield of 93 and 59%,
respectively). The products were characterized by FT-IR and 1D and 2D NMR
spectroscopy, showing that two positions were substituted by tosyl groups: G-6
of the 3-linked β-D-galactosyl unit (G-2) and position C-2 of the 4-linked 3,6-
anyhdro-α-L-galactosyl unit (LA-2). NMR analysis showed a degree of tosylation
of approximately 2. Therefore the chemical tosylation of this important
commercial polysaccharide was successfully achieved, representing a new
agarose backbone for future applications.
Programa de Pós-Graduação em Ciências (Bioquímica)
15 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
EVALUATION OF THE ANTITUMOR ACTIVITY OF NATIVE AND MODIFIED
SULFATED HETERORHAMNANS OBTAINED FROM THE GREEN
SEAWEED GAYRALIA BRASILIENSIS
Ester Mazepa1; Miguel D. Noseda1; Diogo R.B. Ducatti1; Alan G. Gonçalves1;
Rafaela P. Gomes2; Sheila M.B. Winnischofer2; Maria Eugênia D. Noseda1
1Laboratório de Glicobiologia Estrutural de Carboidratos de Algas Marinhas
(GLICAM), Departamento de Bioquímica e Biologia Molecular, Setor de
Ciências Biológicas, Universidade Federal do Paraná, PR, Brasil
2Laboratório de Cultivo Celular e Oxidações Biológicas, Departamento de
Bioquímica e Biologia Molecular, Setor de Ciências Biológicas, Universidade
Federal do Paraná, PR, Brasil, Universidade Federal do Paraná, Paraná, Brasil.
Sulfated polysaccharides produced by seaweeds have great potential for
antitumoral therapy. Study of structure-activity relationship can provide better
understanding of mechanism actions, and enable chemical modifications
enhancing biological effect. The aim of the present study was to evaluate the
effect of the native and chemically modified sulfated polysaccharides isolated
from a seaweed on the viability of cancer cells. A sulfated heterorhamnan (Gb1,
33.6% NaSO3) was obtained by aqueous extraction (80°C) from the green
seaweed Gayralia brasiliensis. Besides rhamnose (63.3 mol%) it contains
xylose (8.8 mol%), galactose (6.4 mol%), glucose (10.1 mol%), and glucuronic
(8.6 mol%) and galacturonic (2.7 mol%) acids. Gb1 was submitted to chemical
modifications: partial depolimerization and oversulfation. The partially
depolymerized product Gb1-S (51.6% NaSO3) contains rhamnose, glucose,
xylose and galactose (91.5, 5.9, 1.1, 0.5 mol%, respectively). Gb1 was
submitted to chemical sulfation giving rise to an oversulfated polysaccharide
(Gb1-OS, 55.6% NaSO3). 1D and 2D NMR analyses of polysaccharides
showed different proportions of 3-linked, 3-linked-2-sulfated, 3-linked-4-sulfated,
3-linked-2,4-sulfated, 2-linked-4-sulfated, 2-linked-3,4-sulfated and 2,3-
disubstituted rhamnose units. The antitumor effect of Gb1, Gb1-S and Gb1-OS
(at 25, 100 and 500 μg/mL) was evaluated against cancer cells (MDA-MB-435)
by MTT assay. Gb1 at 100 and 500 μg/mL reduced cell viability in 23 and 32%,
Gb1-S at 25 μg/mL reduced 22%, and Gb1-OS at 25, 100 and 500 μg/mL
reduced 35, 47 and 48% of cell viability. The oversulfated polysaccharide
showed the highest effect against MDA-MB 435 cells.
Programa de Pós-Graduação em Ciências (Bioquímica)
16 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
EVALUATION OF SIX METHODS FOR TOTAL DNA EXTRACTION FROM
GUT EARTHWORMS FOR MICROBIAL COMMUNITY DIVERSITY AND
METAGENOMICS
Esther Dering Esteves1, George Gardner Brown2, Fábio de Oliveira Pedrosa 1,
Emanuel Maltempi de Souza1, Leda Satie Chubatsu1
1Universidade Federal do Paraná – Department of Biochemistry and Molecular
Biology.
2Embrapa Floresta Colombo.
DNA extraction is a crucial step for many molecular studies including microbial
community diversity in earthworms gut. A variety of methods have been used
for total DNA extraction from earthworms gut, including commercial kits and
manual methods. Extraction methods need to be evaluated for their efficiency,
as DNA degradation and fragmentation during extraction and other effects. In
this work genomic DNA was extracted from three different regions of the
earthworm gut, species Perionyx excavatus, using six different methods,
including commercial kits and manual procedures using SDS or CTAB . The
extracted DNA samples were compared for both yield and DNA quality.
Samples were also tested for efficient amplification of 16S rRNA by PCR.
Earthworm foregut samples had the lower DNA yield independent of the method
used for DNA extraction. Five of the methods had acceptable yields in DNA
extraction for samples from hindgut. All tested methods led to a fragmented
DNA, but one of the manual methods yield a high molecular weight DNA. Only
samples obtained from two of the manual methods and one of the commercial
kits were successful for DNA amplification to all earthworm gut regions. Results
indicate that three of methods could be used for the extraction of total DNA with
the purpose to analyze the microbial biodiversity of earthworm gut. However,
only one of these methods is suitable for metagenomic studies due to the high
molecular weight DNA obtained.
Financial Support: INCT, CAPES, CNPq, PNPD/CAPES
Programa de Pós-Graduação em Ciências (Bioquímica)
17 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
DETERMINATION OF MINIMUM INHIBITORY CONCENTRATION OF
STAPHYLOCOCCUS AUREUS, STAPHYLOCOCCUS EPIDERMIDIS AND
ESCHERICHIA COLI TO DANOFLOXACINO INJECTABLE SOLUTION IN
THE PRESENCE OF THE DEGRADATION PRODUCT OBTAINED UNDER
PHOTOLYTIC CONDITIONS
Everson Willian Fialho Cordeiro¹,²; Renata Medeiros Hilgert¹; Luiz Alcides das
Chagas Batista¹; Cheila Denise Ottonelli Stopiglia², Clésio Sodatelli Paim¹
¹ Laboratório de Pesquisa em Desenvolvimento e Controle de Qualidade de
Medicamentos - Universidade Federal do Pampa, Uruguaiana-RS.
² Laboratório de Microbiologia - Universidade Federal do Pampa, Uruguaiana-
RS.
Danofloxacin is an antimicrobial chemotherapy for exclusive use in veterinary
medicine. The drug has a broad spectrum of action comprising Gram negative
bacteria, Gram positive and also mycoplasmas of clinical interest. The present
study aimed to determine the minimum inhibitory concentration (MIC) of the
drug and evaluating whether the degradation products, quantified by HPLC,
interfere in the activity of the quantitative in vitro assay. The microdilution assay
was performed according to the protocol M100-S25 CLSI (2015). Thus,
Advocin® injectable solution was diluted in Mueller Hinton culture medium and
performed in 96-well plates to give concentrations between 12.8 and 0.03
μg/mL. New tests were performed after the definition of MIC, to assess whether
the drug showed activity changes when it is exposed to forced degradation
conditions. The degradation study showed that the concentration of drug
reduced significantly with time, reaching 74.05% in the first 48 hours of
exposure to light. The results of the microdilution assay demonstrated that S.
aureus and S. epidermidis showed the same MIC (0.12 μg/mL) and E. coli was
more sensitive to the antimicrobial with a MIC of 0.06 μg/mL. However, assays
using the degradation products against S. epidermidis have shown that they
show antibacterial activity, since they reduced the MIC of 0.12 μg/mL to 0.06
μg/mL. Therefore the study demonstrates important contribution to evaluate the
antimicrobial activity and suggests that the degradation products present in vitro
antimicrobial activity.
Programa de Pós-Graduação em Ciências (Bioquímica)
18 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ARAUCARIA ANGUSTIFOLIA CULTURES WITH DIFFERENT EMBRYOGENIC POTENTIAL PRESENT A DISTINCTIVE MITOCHONDRIAL
BIOENERGETICS
Fernando Diego Kaziuk, Ana Luiza D. M. Furlanetto, Andre L.W. Dos Santos,Eny I.S. Floh, Fabiane Fortes, Sílvia M. S. C. Cadena.
Departamento de Bioquímica e Biologia Molecular, Universidade Federal doParaná, Curitiba, Paraná, Brasil.
Departamento de Ciências Biológicas, Universidade Estadual do Paraná,Campus União da Vitória, União da Vitória, Paraná, Brasil.
Laboratório de Biologia Celular de Plantas (BIOCEL), Departamento deBotânica, Instituto de Biociências, Universidade de São Paulo, São Paulo,Brasil.
Araucaria angustifolia (Bert.) O. Kuntze is currently classified in the category CR (species critically endangered) by the IUCN red list of threatened species. Therefore, understanding this plant physiology is essential for its preservation. In this study, it was evaluated hot stress (30ºC) effects on A. angustifolia cell lines with different embryogenic abilities, one cell line being responsive to maturation conditions (SE1cell line) and one cell line that presented blocked development of mature somatic embryos (SE6 cell line). The cells were grown at 25 ± 1°C on semi-solid MSG culture medium (20-21 days of culture) in the dark and submitted to hot stress of 30 ± 1°C for 12h, 24h or 48h. The cells viability evaluated by MTT assay was not affect by all stress conditions. However, the viability of SE6 cells line was higher when compared to SE1 cells lines, submitted or not to stress conditions. As MTT method is based on the activity of mitochondrial dehydrogenases, was also evaluated the oxygen consumption (Oroboros 2k - Oxygraph) in mitochondria isolated from these cells not submitted to stress. Interestingly, the rate of oxygen consumption during states 3 and 4 of respiration and the Respiratory Control Coefficient (RCC) were higher in SE6 cells lines. These results suggest that mitochondrial functions linked to energy provision are stimulated in SE6 cells line and motivate further studies to clarify the involvement of this distinctive characteristic on propagation and maturation of embryogenic Araucaria angustifolia cells.
Financial Support: CNPq, CAPES, INCT-Redoxoma
Programa de Pós-Graduação em Ciências (Bioquímica)
19 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
REDUCTIVE-AMINATION OF Κ-CARRAGEENAN HYDROLYSIS PRODUCTS
Franciely G. Colodi; Miguel D. Noseda; Estela M. Aranha; Mariana M. Carvalho;
Diogo R. B. Ducatti; Maria Eugênia D. Noseda.
Laboratório de Glicobiologia Estrutural de Carboidratos de Algas Marinhas
(GLICAM), Departamento de Bioquímica e Biologia Molecular, Universidade
Federal do Paraná, Curitiba, PR, Brazil.
Carrageenans are sulfated polysaccharides produced by red seaweeds that
present large industrial uses and biological interest. κ-Carrageenan consists of
alternating (1→3)-linked β-D-galactopyranose 4-sulfate and (1→4)-linked 3,6-
anhydro-α-D-galactopyranose units. This negative charged polysaccharide has
been used to assemble films for biomimetic surfaces with multiple functionalities
and to delineate protein delivery systems. The aim of this work was the
functionalization of κ-carrageenan hydrolysate products through reductive-
amination. κ-Carrageenan was obtained from red seaweed Kappaphycus
alvarezzi (aqueous extraction for 4 h at 80 °C), submitted to partial hydrolysis
using TFA (0.1 mol/L) at 65 °C for 15 and 35 min and at 80 °C for 180 min,
giving rise to partially depolymerized products K15, K35 and K180, respectively.
Reductive-amination was carried out in methanol (2.5% w/v) with propane-1,3-
diamine (2 eq., pH 5.5) and sodium cyanoborohydride (2.5 eq.). The mixtures
were stirred for 15 h at 55 °C, giving aminated fractions K15a, K35a and K180a.
κ-Carrageenan hydrolysis products K180, K35 and K15 are, respectively,
composed mainly by κ-carrabiose, κ-carratetraose and κ-oligosaccharides
larger than six units. HSQC NMR spectrum of K15a showed signals
corresponding to aminated 3,6-anhydro-α-D-galactopyranosyl units (C-1/H-1 at
57.1/3.50 ppm and C-2/H-2 at 69.1/3.98 ppm). Correlations at 50.0/3.32, 3.35;
32.8/2.78 and 45.0/3.24 ppm were attributed to -CH2- of N-linked propane-1,3-
diamine. Therefore, functionalized k-amino-oligosaccharides with different
degrees of polymerization can be obtained by reductive-amination, providing an
interesting approach to develop novel biomaterials.
Programa de Pós-Graduação em Ciências (Bioquímica)
20 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
SELECTIVE C-6 OXIDATION OF CARRAGEENANS AND EVALUATION OF
THE ANTICOAGULANT PROPERTIES
Gislaine C. dos Santos1, Diogo R. B. Ducatti1, Miguel D. Noseda1, Alan G.
Gonçalves2
1Departamento de Bioquímica, Universidade Federal do Paraná, Curitiba, Brasil
2Departamento de Farmácia, Universidade Federal do Paraná, Curitiba, Brasil
Carrageenans are polysaccharides extracted from extracellular matrix of red
seaweeds that belong to the family of sulfated galactans. They have a broad
spectrum of biological activities, including anticoagulant properties, which are
related to the sulfation pattern, molecular weight and chain conformations.
Chemical modifications of polysaccharides have been reported in order to
improve those properties. The selective C-6 oxidation of polysaccharides can
produce polyuronic acids derivatives with new biological and physical
properties, for example, the gelation, complexation, adhesion as well as a
variety of biological activities such as anticoagulant activity. Selective oxidative
methods using 2,2,6,6-tetramethylpiperidine-1-oxyl free radical (TEMPO) as
catalyst in water have been reported in the literature for several soluble and
insoluble polysaccharides such as starch, maltodextrin, cellulose,
galactomannan, glucan, hyaluronic acid, chitin, chitosan and xanthan in order to
generate polymers with potential biological and biotechnological applications.
Thus, the aim of this work was the selective C-6 oxidation of β-D-Galp units in
kappa-, iota-, iota/nu-, theta e lambda-carrageenan using TEMPO as catalyst
and trichloroisocyanuric acid (TCCA) in a carbonate buffer (pH 9.6). All the
samples were characterized by NMR (1D and 2D). The anticoagulant activity of
native and oxidized fractions was examined by aPTT in vitro.
Financial Support: CAPES, CNPq, Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
21 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
SCALE-UP OF BIODIESEL SYNTHESIS IN FIXED BED REACTORS USING
THE FERMENTED SOLID BY BURKHOLDERIA LATA
Dias, Glauco Silva1; Luz Jr., Luiz Fernando de Lima2; Krieger, Nadia3; Mitchell,
David Alexander1
1Laboratório de Tecnologia Enzimática e Biocatálise, Dep. de Bioquímica e
Biologia Molecular, UFPR, PR;
2Dep. de Engenharia Química, UFPR; (3) Dep. de Química, UFPR de
Farmácia, Universidade Federal do Paraná, Curitiba, Brasil
The biodiesel is produced industrially by homogeneous alkaline
transesterification of the triacylglycerols of vegetable oils with methanol. This
process requires steps for the removal from the biodiesel of the catalyst and of
the salt that is formed, for the treatment of the alkaline effluent that is generated
and for the recovery of the glycerol by product that is produced. One possible
strategy for avoiding problems associated with chemical catalysis is to use
lipases as catalysts. Recently, in our laboratory, a process was developed with
the intention of reducing the costs of the enzymatic route. The objective of the
current work was to increase the scale of this biodiesel production process. To
this end, a bioreactor consisting of three packed-beds in series was filled with a
total of 120 g of fermented solid, produced using Burkholderia lata and 1245 g
of a medium comprised of olein and ethanol was recirculated through the
reactor from a reservoir. The best results being a giving 88% conversion in 24
h. This system was operated for six consecutive 48-h cycles. In the first cycle, 1
kg of esters was produced, while the six cycles produced a cumulative total of
4.7 kg of esters, which is equivalent to 39 g of ester produced per g of
fermented solid that was used. These results are promising, since they
demonstrate that high yields can be maintained in the scale-up of enzymatic
biodiesel production processes involving fermented solids as the catalyst.
Programa de Pós-Graduação em Ciências (Bioquímica)
22 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
MONOCLONAL ANTIBODIES DEVELOPMENT FOR ADAM33 AND ITS
CLINICAL APPLICATION
Graciele C. M. Manica1, Marco A. S. de Oliveira2, Andressa Chequin1, Edneia
A. S. Ramos1, Liliane M. B. Klassen1, Mariana Busato Toledo1, Yara de Oliveira
Brandão1, Isis Venturi Biembengut1, Emanuel M. De Souza2, Giseli Klassen1
1 Dep. de Patologia Básica, Laboratório de Epigenética;
2 Dep. de Bioquímica e Biologia Molecular, UFPR, PR, Brasil
The biodiesel is produced industrially by homogeneous alkaline
transesterification of the triacylglycerols of vegetable oils with methanol. This
process requires steps for the removal from the biodiesel of the catalyst and of
the saltADAM33 gene is down-regulated in human breast cancer by promoter
hipermethylation. In order to make a clinical usefull tool, a recombinant
truncated ADAM33 protein containing the cystein rich domain was used to
immunize Balb/c mice and monoclonal antibody anti-ADAM33 was produced
(GMGK06). The specificity of the antibody was tested by western blot (WB) and
Immunocytochemical (IHQ). The WB results showed that GMGK06 has high
specificity for ADAM33 protein since no signal was detected in the MDA-MB-
231, a cell line ADAM33 negative. However, the ADAM33 expressing breast cell
lines MCF7 and PMC42, showed a single signal of approximately 37 kDa. The
antibody was used to test 44 cases of human breast cancer by
immunohistochemical assay. We found three different scores of ADAM33 in
breast cancer samples: 2 (weak), 3 (intermediate) and 4 (strong). Anti-ADAM33
antibodies may be useful in further studies to understand the biological
mechanisms of the breast cancer, furthermore, to evaluate the function of
ADAM33 in normal physiological states, different types of cancer and other
diseases.
Programa de Pós-Graduação em Ciências (Bioquímica)
23 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ANTICOAGULANT AND ANTITHROMBOTIC ACTIVITY OF NATIVE AND
PARTIALLY DEGRADED GLYCOGLUCURONOMANNAN FROM VOCHYSIA
THYRSOIDEA AFTER CHEMICAL SULFATION
Helyn Priscila de Oliveira Barddal, Ana Helena Pereira Gracher, Fernanda
Fogagnoli Simas-Tosin, Marcello Iacomini, Thales Ricardo Cipriani
Dep. de Bioquímica e Biologia Molecular, UFPR, PR, Brasil
Heparin has great clinical importance as anticoagulant and antithrombotic
agent. However, because of its risks of causing bleeding and contamination by
animal pathogens, several studies aim to obtain alternatives to heparin. In the
search for anticoagulant and antithrombotic agents from a non-animal source, a
glycoglucuronomannan from the gum exudate of the plant Vochysia thyrsoidea
was partially hydrolyzed, and both native and partially degraded
polysaccharides were chemically sulfated, yielding VThS and Ph-VThS
respectively. Methylation analysis indicated that sulfation occurred preferentially
atthe O-5 position of arabinose units in the VThS and at the O-6 position of
mannose units in Ph-VThS. In vitro aPTT assay showed that VThS and Ph-
VThS have anticoagulant activity, which could be controlled by protamine, and
ex vivo aPTT assay demonstrated that Ph-VThS is absorbed by subcutaneous
route. Like heparin, they were able to inhibit α-thrombin and factor Xa by a
serpin-dependent mechanism. In vivo, VThS and Ph-VThS reduced thrombus
formation by approximately 50% at a dose of 40 IU/kg, similarly to heparin. The
results demonstrated that the chemically sulfated polysaccharides are
promising anticoagulant and antithrombotic agents.
Financial Support: CAPES, CNPq, UFPR, Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
24 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
THE EFFECTS OF XILOGLUCANA EXTRACTED FROM COPAIFERA
LANGSDORFFII (XGC) AND ITS COMPLEX WITH OXOVANADIUM IV/V
(XGC:VO) ON MITOCHONDRIAL BIOENERGETICS
Fernandes, K.L.M¹; Petkowicz, C. L. O¹; Noleto, G.R¹; Cadena, S.M.S.C¹
¹ Department of Biochemistry and Molecular Biology - Universidade Federal do
Paraná (UFPR), Paraná, Brasil.
It was shown that xiloglucana extracted from Copaifera langsdorffii (XGC) and
its complex with oxovanadium (XGC:VO) were cytotoxic to B16F10 murine
melanoma cells. This effect was related to the impairment of cells respiration. In
this study we evaluated the effects of these polymers on respiration parameters,
using glutamate plus malate as oxidizable substrates, in isolated rat liver
mitochondria. The activities of NADH and succinate oxidases in disrupted
organelle were also evaluated. XGC (0.5-25 μg.mL-1), did not affect the
respiration in intact organelle; however, the polysaccharide reduced the oxygen
consumption in uncoupled mitochondria (0.5 µM of FCCP) by ~30%.
Interestingly, XGC:VO reduced by 13% (0.5-25 μg.mL-1) the respiratory rate of
the state 3 but did not affect the state 4. The inhibition of state 3 resulted in a
decrease also by ~13% of Respiratory Control Coefficient (RCC) while ADP/O
ratio was unchanged. On the uncoupled state, the decrease of oxygen
consumption by XGC:VO was more pronounced (~32%). The activity of NADH
oxidase evaluated by oxygen consumption was not affected by both XGC and
XGC:VO. However, succinate oxidase activity was increased in the presence by
XGC in ~90% already at lower concentrations but, the enzyme activity was
unchanged by XGC:VO. These results suggest that the presence of vanadium
was essential for the inhibition of respiration in intact mitochondria oxidizing
substrates of complex I (glutamate plus malate). On the other hand, the
stimulation of succinate oxidase only for native XGC suggests that this effect is
abolished by the metal presence.
Programa de Pós-Graduação em Ciências (Bioquímica)
25 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
STRUCTURAL CHARACTERIZATION AND STUDY OF THE ANTI-
INFLAMMATORY POTENTIAL OF THE POLYSACCHARIDES OF
CABERNET FRANC, CABERNET SAUVIGNON AND SAUVIGNON BLANC
WINES
Iglesias de Lacerda Bezerra, Adriana Rute Cordeiro Caillot, Arquimedes Paixão
Santana Filho, Guilherme Lanzi Sassaki
Department of Biochemistry and Molecular Biology - Universidade Federal do
Paraná (UFPR), Paraná, Brasil.
Introduction: There are few works about characterization of polysaccharides of
wines. The structure and amounts of polysaccharides released depend on the
wine-making process and can influence the sensory properties and quality of
the wines. This work aimed structural characterization of the polysaccharides
found in three types of wines: Cabernet Franc (WCF), Cabernet Sauvignon
(WCS) and Sauvignon Blanc (WSB). Material and methods: The wines were
concentrated and the polysaccharides were obtained via ethanolic precipitation
followed centrifugation, dialysis and freeze dry. The polysaccharides were
analyzed by nuclear magnetic resonance. Monosaccharide composition was
determined after total hydrolysis with TFA 100°C/14h by quantitative-HSQC.
Homogeneity analyses were performed by HPSEC-MALLS. The anti-
inflammatory potential of the polysaccharides through inhibition of NF-Kβ in
Raw blue cells Results: Polysaccharide yields were: 1.5% (WCF), 0.5% (WCS)
and 0.2% (WSB). WCF showed Gal-(32.1%), Ara-(31.4%), Rha-(9.1%), GalA-
(9.2%), Glc-(10.6%) and Man-(6.3%); WCS showed Gal-(18.6%), Ara-(15.1%),
Rha-(9.2%), GalA-(12.0%), Glc-(18.7% ) and Man-(26.4%); and WSB showed
Gal-(19.1%), Ara-(19.2%), Rha-(4.7%), GalA-(4.1%), Glc-(12.7% ) and Man-
(40.2%). The total uronic acids content was determined for WCF, WCS and
WSB, giving rise to 15.2%, 16.4% and 8.6%, respectively. All the samples
showed a heterogeneous elution profile, suggesting the presence of
polysaccharide mixture. HSQC-NMR spectroscopy indicated the presence at
least four polysaccharides in all samples: An arabinogalactan type II, type I
rhamnogalacturanan, dextrin, and mannan. All the simples inhibited of NF-Kβ in
vitro. Conclusion: The results suggest that the HSQC NMR of the
polysaccharides can furnish a fingerprint for each wine, since the profile of the
mixtures had different yields and quantities, aiding for a non-volatile based
singular signature. As also, the simples showed anti-inflammatory potential, but
they will still be made other experiments to prove these results.
Programa de Pós-Graduação em Ciências (Bioquímica)
26 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
INTRAGENIC DNA METHYLATION REGULATES MMP9 GENE
EXPRESSION IN BREAST CANCER
Liliane M. B. Klassen1, Graciele C. M. Manica1, Isis V. Biembengut1, Edneia A.
S. Ramos1, Andressa Chequin1, Isis, Mariana B. Toledo1, Yara de O Brandão1,
Emanuel M. de Souza2, Giseli Klassen1
1Department of Basic Pathology, Federal University ofParana.
2Department of Biochemistry and Molecular Biology, Federal University of
Parana.
Breast cancer is the most frequently diagnosed cancer and the leading cause of
cancer death among women worldwide. Metastasis remains a major challenge
for the clinical management and prognosis of patients with cancer. The
metalloprotease MMP-9 play a critical role in the first step of metastasis through
extracellular matrix degradation. In cancer from breast, cervical, prostate and
many others MMP-9 have been correlated with poor prognosis. The MMP9
gene could be regulated by epigenetic mechanisms such as histone
modifications however DNA methylation remains to be further clarified. This
study starts with an in silico study of MMP9 gene showed two intragenic CpG
islands. Our goal was verify the effect of these DNA region in MMP9 gene
expression. The quantification in breast cancer cell lines with or without
decitabine (DAC) treatment showed that MCF7 and MDAMB436 expressed
MMP9 only after demethylating agent treatment. The sequencing data obtained
from the two DNA regions with high CpG content (CpG island) showed one
specific sequence with 273 pb between CpGs 11 and 29 in the second CpG
island differentially methylated. This specific region was studied in breast
cancer samples that reveal the similar results with demethylation in positive
MMP-9 samples. Taken together these results showed a new possible
mechanism of DNA methylation and gene expression regulation since no
description of intragenic DNA region was showed until now.
Programa de Pós-Graduação em Ciências (Bioquímica)
27 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
MICROPARTICLES OF CACAO POD HUSKS PECTIN AND 5-ASA: A
PRELIMINARY INVESTIGATION
Vriesmann, L.C.1; Franco, C.R.C.2; Lucinda-Silva, R.M.3 Petkowicz, C.L.O.1
1Departamento de Bioquímica e Biologia Molecular, UFPR, Curitiba-PR, Brazil
1Departamento de Biologia Celular, UFPR, Curitiba-PR, Brazil2
3Departamento de Ciências Farmacêuticas, UNIVALI, Itajaí-SC, Brazil
Pectins are polymers from plant cell wall widely used as gelling and stabilizing
agents in the food industry. Recently, they are suggested to the development of
colon-specific therapeutic systems. In this work, we employed a highly
acetylated HM pectin (OP) from cacao pod husks to prepare microparticles
containing 5-ASA, a drug employed in inflammatory bowel disease. The
microparticles were obtained by spray drying, using solutions of pectin and 5-
ASA in the proportions 1:1; 2:1; 3:1 and 4:1. Sample 1:1 has the higher yield
(68%) and 86% of drug incorporation efficiency. This efficiency was improved
when the proportion of pectin increased, with sample 4:1 reaching 98%
efficiency. From MEV, the microparticles presented <5 micrometers and an
irregular surface, more pronounced in sample 1:1. Liberation assays were
performed simulating the gastrointestinal transit. Although more investigation is
necessary to improve the drug protection in the gastric pH, the obtained results
indicate that this pectin showed better tendency of drug retention when its
content is increased, being a promissory polymer to be studied as an adjuvant
in systems containing drugs directed to colon liberation or action.
Programa de Pós-Graduação em Ciências (Bioquímica)
28 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
UNRAVELLING THE TRANSCRIPTIONAL REGULATORY NETWORK OF
THREE FNR PROTEINS FROM HERBASPIRILLUM SEROPEDICAE SMR1
USING CHIP-SEQ AND RNA-SEQ
Marcelo Bueno Batista1,2, Govind Chandra2, Emanuel Maltempi de Souza1,
Rose Adele Monteiro1, Ray Dixon2
1Department of Biochemistry and Molecular Biology, Universidade Federal do
Parana, P.O. Box 19046, Curitiba, PR 81531-990, Brazil
2Department of Molecular Microbiology, John Innes Centre, Colney Lane,
Norwich NR4 7UH, UK
H. seropedicae SmR1 is an endophytic aerobic bacterium capable of fixing
atmospheric nitrogen under conditions of nitrogen and oxygen limitation. To
efficiently adapt to low O2 levels, H. seropedicae, as in the case of other
bacteria, takes advantage of a branched respiratory chain comprising different
types of terminal oxidases, including oxidases predicted to have high affinity for
oxygen. Remarkably this organism has genes coding for three Fnr proteins,
designated as Fnr1, Fnr2 and Fnr3. Using genome-wide transcriptional
profiling in combination with physiological characterisation, we previously
observed that efficient reconfiguration of the respiratory chain in H. seropedicae
under low oxygen availability relies on transcriptional regulation of gene
expression by these Fnr proteins. However, we were not able to define the
specific regulons and functional roles of each of the three Fnr transcription
factors. In this study we have used a combination of RNA-Seq transcriptional
profiling of single fnr mutants together with a ChIP-Seq approach to address the
functions of three Fnr orthologs encoded by H. seropedicae genome. Although
Fnr1 and Fnr3 regulate discrete classes of genes, we have identified another
group of genes that are jointly regulated by both transcription factors.
Promoters in this class are bound by both Fnr1 and Fnr3, potentially indicative
of regulation by Fnr1-Fnr3 heterodimers. In contrast Fnr2 is apparently an
oxygen insensitive protein responsible for the transcriptional activation of the
bo3-type respiratory oxidase.
Programa de Pós-Graduação em Ciências (Bioquímica)
29 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
IDENTIFICATION OF AN EXOGLUCANASE OBTAINED FROM
METAGENOMIC LIBRARY OF BRAZILIAN ATLANTIC FOREST SOIL
Marcelo Scarduelli, Bruno Afonso Ramos Cassilha, Helisson Faoro, Fábio de
Oliveira Pedrosa, Luciano Fernandes Huergo e Emanuel Maltempi de Souza
Departamento de Bioquímica e Biologia Molecular- Universidade Federal do
Paraná
Cellulases have many industrials uses, from polishing fabrics to biofuel
production, especially second generation bioethanol. The group of cellulases
include endoglucanases, exoglucanases and beta-glucosidases. Searching for
new cellulases, prospection in metagenomic libraries could lead to discover
novel enzymes. This technique allows to capture all the genetic material in a
sample, covering nucleotidic sequences both cultivable and uncultivable
species in laboratory conditions. This study aimed to identify a
celulase/exoglucanase in metagenomic library of Brazilian Atlantic Forest soil.
This library was previously constructed. Sequence comparison was used for
identification of cellulases. Selected genes were cloned and overexpressed in
E. coli. Purification was performed by Ni2+ affinity chromatography (HiTrap
Chelating, GE Healthcare). The enzymatic activities were calculated with DNS
assay (3 5-dinitrosalicylic acid). After comparison analysis, one gene was
identified as exoglucanase, with 66 kDa and conserved domain for family 9
glycosyl hydrolases. This gene showed low identity with submitted genes: 70%
of identity with a hypothetical protein from Segetibacter koreensis or 66% with
glycosyl hydrolase from Chitinophaga pinensis. Preliminary results showed
enzymatic activity only in high level of NaCl (1 and 2 M), when using
regenerated amorphous cellulose (RAC) as substrate. Others enzymatic assays
will be made in order to quantify the cellulase activity and confirm the
halotolerance of this exoglucanase.
Programa de Pós-Graduação em Ciências (Bioquímica)
30 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
PERIODATE OXIDATION OF ULVANS FOR INTRODUCTION OF NEW
REACTIVE GROUPS
Mariana M. de Carvalho1, Miguel D. Noseda1, Franciely G. Colodi1, Rilton A. de
Freitas2, Maria Eugênia D. Noseda1
1Laboratório de Glicobiologia Estrutural de Carboidratos de Algas Marinhas
(GLICAM), Departamento de Bioquímica e Biologia Molecular, Universidade
Federal do Paraná, Paraná, Brasil.
2Biopol – Departamento de Química, Universidade Federal do Paraná, Paraná,
Brasil.
Green seaweeds from Ulva spp. biosynthesize sulphated polysaccharides
named ulvan. Considering they are still unexplored, chemical modifications may
increase their possible applications. The aim of this study was to introduce
aldehyde groups in ulvans by periodate oxidation, originating new reactive
groups for chemical modifications. Ulvans from Ulva fasciata were obtained by
aqueous extraction (80 ºC) resulting in F2 (20.9% SO3Na, 16.0% uronic acids),
which according to monosaccharide composition and NMR analyses (13C, 1H
and HSQC), was built up of the following diads: [→4)-β-D-glcpA-(1→4)-α-L-
rhap3S-(1→], [→4)-β-D-xylp-(1→4)-α-L-rhap3S-(1→], [→4)-β-D-xylp2S-(1→4)-
α-L-rhap3S-(1→] and ([→4)-α-L-idopA-(1→4)-α-L-rhap3S-(1→]). F2 was
submitted to periodate oxidation for 24, 48 and 72 h resulting in the partially
oxidized polysaccharide fractions F2-A, F2-A1 and F2-A2 respectively.
Oxidation was confirmed by FTIR (aldehyde bands at 1750 cm-1) and 13C
NMR (hemicetals at 90-91 ppm). Moreover in the 13C spectra, the decrease in
the C1 signals of [→4)-β-D-xylp-(1→] (104.8 ppm) and ([→4)-α-L-idopA-(1→]
(104.1 ppm) indicates that these units were, as expected, oxidized. Units with a
lack of vicinal hydroxyls such as ([→4)-β-D-xylp2S-(1→] and [→4)-α-L-rhap3S-
(1→) were not oxidized. Noteworthy, the units of [→4)-β-D-glcpA-(1→] were not
oxidized, although they contain vicinal hydroxyls. This may occur when the unit
adopts a conformation that prevents periodate oxidation or by the formation of
hydrogen bonds between COOH at C6 and OH at C3/C4. These results indicate
that the introduction of aldehydes groups was successful, giving rise to
aldheydic ulvans. Chemical modifications from periodate oxidized ulvans are in
progress. Physical-chemical properties and biological activities of these new
polysaccharides are under study.
Financial Support: CAPES, CNPq, Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
31 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
NTRYX AND NTRBC SYSTEMS ARE REQUIRED TO INDUCE GENES
ENCODING THE ASSIMILATORY NITRATE REDUCTASE IN
HERBASPIRILLUM SEROPEDICAE SMR1
Bonato, P.1; Tadra-Sfeir, M. Z.1; Camilios-Neto, D.2; Wassem, R.1; Rigo, L.u.1;
Pedrosa, F. O.1; Souza, E.M.1; Chubatsu, L.S.1
1Núcleo de Fixação de Nitrogênio, Departamento de Bioquímica e Biologia
Molecular, Universidade Federal do Paraná;
2Departamento de Bioquímica e Biotecnologia, Universidade Estadual de
Londrina
Nitrate is the main inorganic source of nitrogen found in soil. The diazotrophic
and plant-associative Herbaspirillum seropedicae SmR1 has genes encoding
for two nitrate reductases: the assimilatory (NAS) and the respiratory nitrate
reductases (NAR). The ntrY and ntrC mutant strains do not grow on nitrate as
the only nitrogen source suggesting that the NtrYX and NtrBC systems are
required to induce genes important for nitrate assimilation. Given that, H.
seropedicae wild type, and ntrY and ntrC mutant strains were cultivated
aerobically in low nitrogen (1 mM NH4Cl) until cells reached OD600 0.4, then
cells were divided in three parts: one stored in RNAlater and the other two
incubated for more 30 minutes after addition of 10 mM KNO3 or 10 mM NH4Cl.
Samples from the three bacterial strains in these three different conditions were
submitted to RNA extraction, cDNA libraries construction and DNA sequencing
using an Ion Proton Sequencer. Reads were mapped against the H.
seropedicae genome using CLC Genomics Workbench 7.0 and differentially
expressed genes were analysed using DESeq 2.0. The
narKnirBDHSERO_RS14545nasA operon, which encodes the NAS system,
showed the highest induction rate after nitrate addition in the wild type strain,
when compared to ammonium or to low nitrogen conditions. In contrast these
genes were downregulated in both ntrY and ntrC mutant strains cultivated with
nitrate. These results were validated with lacZ reporter fusions. Together these
results indicate that NtrYX and NtrBC systems are required to activate the
narKnirBDHSERO_RS14545nasA operon.
Programa de Pós-Graduação em Ciências (Bioquímica)
32 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
MODIFICATION ON THE 2’-DEOXYGUANOSINE OXIDATION BY SINGLET
MOLECULAR OXYGEN PATHWAY BY GLUTATHIONE
Patrícia S. Peres1, Andressa Valerio1, Silvia M. S. C. Cadena1, Sheila M. B.
Winnischofer1, Alexsandra C. Scalfo2, Paolo Di Mascio2, Glaucia R. Martinez1
1Laboratório de Oxidações Biológicas, Departamento de Bioquímica e Biologia
Molecular, Setor de Ciências Biológicas, Universidade Federal do Paraná
(UFPR), Curitiba, PR, Brazil.
2Departamento de Bioquímica, Instituto de Química, Universidade de São
Paulo, São Paulo,SP,Brazil.
The oxidation of the free nucleoside 2’-deoxyguanosine (dGuo) by singlet
molecular oxygen (1O2) has been studied over the three last decades due to the
major role of DNA oxidation products in process such as ageing, mutation and
carcinogenesis. In the present work we investigated the dGuo oxidation by 1O2
in the presence of the important low molecular antioxidant, glutathione, in its
reduced (GSH) and oxidized (GSSG) forms. There were applied different
conditions of concentration, pH, time of incubation, and the use of a [18O]-
labeled thermolabile endoperoxide naphthalene derivative as a source of [18O]-
labeled 1O2. Data was obtained through high performance liquid
chromatography (HPLC) and HPLC coupled to micrOTOFQ-II analysis of the
main oxidation products: the diastereomers of spiroiminodihydantoin-2′-
deoxyribonucleosides (dSp) and 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-
oxodGuo). An intriguing result was that 8-oxodGuo levels increased by 100 fold
when dGuo was oxidized by 1O2 in the presence of GSH and by 2 fold in the
presence of GSSG, while dSp levels dropped to zero for both conditions. All
data from dGuo, 8-oxodGuo and dSp quantification together with the analysis of
residual GSH/GSSG content in each sample strongly suggest that glutathione
modifies the mechanism of dGuo oxidation by 1O2 by disfavoring the pathway of
dSp formation.
Programa de Pós-Graduação em Ciências (Bioquímica)
33 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
INFLUENCE OF MOLAR MASS AND CONCENTRATION ON THE
THERMOGELATION AND INTERFACIAL PROPERTIES OF
METHYLCELLULOSES
Pauline L. Nasatto 1,2, Joana L. M. Silveira1, Frédéric Pignon2, Marguerite
Rinaudo3, Miguel D. Noseda1, Maria Eugênia R. Duarte1.
1Laboratório de Glicobiologia Estrutural de Carboidratos de Algas Marinhas,
Departamento de Bioquímica e Biologia Molecular, Federal University of
Paraná, Curitiba, Paraná, Brazil.
2Laboratoire Réologie et Procedes, Université Grenoble Alpes, Grenoble,
France.
3Biomaterials Applications, Grenoble, France.
Four methylcelluloses having the same average degree of substitution and
distribution of methyl groups, but different molar masses were investigated, their
thermogelation correlation with the molar mass and concentration in aqueous
medium, as well their interfacial interactions, at room temperature and at very
low polymer concentrations were studied. Heating process was specially
studied to analyse the two steps of gelation using rheometry, a large hysteresis
between heating and cooling ramps was observed whatever the conditions. At
low temperature, in the sol state, viscosity depends on the concentration and
molar mass. Over 30 °C a gel like behaviours was observed including two steps
(the second step is a strong gel with phase separation) having storage moduli
which are nearly independent of polymer molar mass but directly related to
polymer concentration. The surface tension (σ) at the water/air interface was
determined for the progressive addition of methylcellulose up to 100 mg/L; σ
starts to decrease over 1 mg/L up to the critical aggregation concentration
(CAC) at 10 mg/L. Curves describing the influence of polymer concentration on
σ are independent of the molar mass at equilibrium. The adsorption of
methylcellulose on silica particles was estimated from ζ-potential
measurements. Those results were interpreted in terms of an increase of the
adsorbed layer thickness at the interface when the molar mass of
methylcellulose increases. It was demonstrated that methylcellulose is
adsorbed, forming trains and loops, at the interface based on the equilibrium
between surface free energy and solvent quality.
Programa de Pós-Graduação em Ciências (Bioquímica)
34 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
EVALUATION OF CYTOTOXICITY OF ZINC PHTHALOCYANINE LOADED
IN SUPERPARAMAGNETIC POLY(METHYL METHACRYLATE)
NANOPARTÍCLES VIA PHOTODYNAMIC THERAPY
Paulo Emilio Feuser1, Eduardo Ricci-Júnior2, Claudia Sayer1and Pedro
Henrique H. de Araújo1
1Department of Chemical Engineering and Food Engineering, Federal
University of Santa Catarina, Brazil
2Faculty of Pharmacy, Federal University of Rio Janeiro, Brazil
Superparamagnetic nanoparticles (MNPs) are promising materials for
hyperthermia treatment and magnetic targeting systems. PDT is a current
therapy that involves the administration of a non-toxic dye and the activation of
photosensitizers with visible light. The aim of this work was the simultaneous
encapsulation of magnetic nanoparticles (MNPs) and zinc (II) phthalocyanine
(ZnPc) in poly(methyl methacrylate) (PMMA) nanoparticles (NPs) by
miniemulsion polymerization and to evaluate the photobiological activity against
murine fibroblast (L929) and human lung adenocarcinoma epithelial cells
(A549). NPs presented an average diameter of 104 ±2.5 nm with a
polydispersity index (PdI) of 0.14 ±0.03 and negative surface charge - 47 ± 2.2
mV (pH 7.4 ±0.1). The release of ZnPc from PMMA NPs was slow and
sustained without the presence of burst effect, indicating a homogeneous
distribution of the drug in the polymeric matrix. NPs did not present any
cytotoxicity effect on L929 cells, after activation with visible light at 675 nm.
However, NPs showed considerable cytotoxic effect on A549 cells only after
activation with visible light at 675 nm photodynamic therapy (PDT). The
preparation of ZnPc loaded in superparamagnetic PMMA NPs with sustained
release can be a new alternative for cancer treatment leading to significant
tumor regression after minimum doses light with a targeted drug delivery.
Programa de Pós-Graduação em Ciências (Bioquímica)
35 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
EVALUATION OF B16-F10 CELL PROLIFERATION UNDER
MELANOGENESIS STIMULUS IN THE PRESENCE OF ANTIOXIDANT N-
ACETYLCYSTEINE
Paulo Szwarc, Tassiele A. Heinrich, and Glaucia R. Martinez
Departamento de Bioquímica e Biologia Molecular, Setor de Ciências
Biológicas, Universidade Federal do Paraná, Curitiba, PR, Brazil
Melanoma is a skin tumor with high pigmentation (melanin). Previous results by
our group have shown that B16-F10 cells (murine melanoma model) with
induced melanogenesis feature increased levels of reactive oxigen species
(ROS) and changes in metabolism, citoskeleton and celular resistance proteins,
cell cycle halt at G1 and fluroescent marking with Pyronin-Y, suggesting that
induced melanogenesis promotes entry into a quiescent state as a protection
mechanism against increased melanin production. This study's main objective
was to evaluate cell proliferation on B16-F10 cells in the presence of antioxidant
N-acetylcysteine (NAC), aiming to better understand the role that ROS plays in
relation to melanogenesis-induced decreased cell proliferation. The cells were
cultured, plated and treated with RPMI 1640 medium supplemented with L-
tyrosine and NH4Cl to induce melanogenesis, and also received NAC
treatment. Cell count was done using an hemocytometer between 12 h intervals
for 48 h, and the results were plotted in a cell growth graph. A decrease in
melanogenesis-induced cell proliferation was observed in comparison to non-
induced cells, and melanogenesis-induced cells that received NAC treatment
had higher numbers of cells than the control. This highlight a possible
involvement of ROS in melanogenesis-induced decreased cell proliferation.
Programa de Pós-Graduação em Ciências (Bioquímica)
36 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
STRUCTURAL CHARACTERIZATION OF A TYPE II ARABINOGALACTAN
FROM CHAMOMILLA RECUTITA [L.]RAUSCHERT INFUSION
Chaves, Pedro Felipe Pereira; Iacomini, Marcello; Cordeiro, Lucimara Mach
Côrtes
Biochemistry and Molecular Biology Department of Federal University of Paraná
Chamomile (Chamomilla recutita [L.] Rauschert) is one of most commonly used
species in phytotherapy and is included in the pharmacopoeia of almost all
countries. Some of the known pharmacological effects are attributed to the
presence of secondary metabolites, but it is not known whether other molecules
such as polysaccharides, are working together to these effects. The chemical
structure of polysaccharides have direct relationship with the biological activities
and elucidate this characteristics is of paramount importance. There are few
studies on the chemical structure of chamomile polysaccharides. Therefore, the
present study aimed to structurally characterize a type II arabinogactan
obtained from chamomile infusion. After a series of ultrafiltrations, a Fehling
precipitation, a enzyme treatment and a elution in chromatographic column a
fraction was obtained and analyzed by monosaccharide composition,
methylation and HSQC/DEPT. The mainly monosaccharides observed were
arabinose and galactose. The major methylated derivative observed was
corresponding of 3,6-O-substituted galactose units. Other methylated
derivatives observed, were corresponding to galactose (terminal 6-O, 3-O and
2,3,6-tri-O-substituted), to arabinose (terminal 5-O- and 3,5-di-O-substituted)
and terminal glucuronic acid units. The HSQC/DEPT spectrum showed signals
assigned to the C1/H1 of α-L-Araf-(1→, →5)-α-L-Araf-(1→, β-D-Galp-(1→, →3)-
β-D-Galp-(1→/→3,6)-β-D-Galp-(1→ e →6)-β-D-Galp-(1→ units, to C6/H6
substituted of the β-D-Galp units, to C5/H5 substituted of the α-L-Araf units and
to free C6/H6 and C5/H5 of the α-L-Araf-(1→, β-D-Galp-(1→ and →3)-β-D-
Galp-(1 units. In conclusion, a type II arabinogalactan that the main chain
consists of (1 → 3)-linked galactose units 6-O-substituted by side chains was
isolated from the chamomile infusion.
Programa de Pós-Graduação em Ciências (Bioquímica)
37 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
STEADY AND DYNAMIC SHEAR RHEOLOGICAL PROPERTIES
OFGABIROBA PULP
(CAMPOMANESIA XANTHOCARPA BERG)
Barbieri, Shayla F. 1; Petkowicz, Carmen L. O. 1; Ruthes, Andrea C.2; De
Godoy, Rossana C. B.3; Silveira, Joana L. M. 1
1Biochemistry and Molecular Biology Department, Federal University of Paraná,
CEP 81.531-980, Curitiba-PR, Brazil
2Division of Glycoscience, Royal Institute of Technology - KTH, Sweden
3Empresa Brasileira de Pesquisa Agropecuária - Embrapa Florestas, CEP
83.411-000, Colombo-PR, Brazil
Gabiroba (Campomanesia xanthocarpa Berg) is a Brazilian native fruit. Its pulp
presented 82 ± 0.8% of moisture content, while different polysaccharides:
pectin, hemicellulose and cellulose compose 17 ± 0.8% of dry weight.
Monosaccharide composition of pectin fractions showed mainly arabinose (Ara
40-60%) and galacturonic acid (GalA 20-42%). The rheological properties of
gabiroba pulp were evaluated by steady-state shear experiments where pulp
exhibited a non-Newtonian pseudoplastic behavior and also showed a yield
stress minimum to initiate the flow related to the material’s internal structure
which must be broken. The presence of a yield stress is a typical characteristic
of multiphase materials as fruit pulps and juices, which are formed by a
dispersion of insoluble components. In dynamic rheological analysis, the
gabiroba pulp presents gel behavior (frequencies 0.01-100 Hz, at 25°C).
Thermal stability as a gel behavior was observed for the gabiroba pulp at
temperatures from 5-95°C, 1 Hz. This stability is suitable for use of the pulp in
food formulations, such as the production of jelly.
Financial Support: Rede Nanoglicobiotecnologia MCT/CNPQ, Pronex
Carboidratos, CNPq
Programa de Pós-Graduação em Ciências (Bioquímica)
38 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
CONSTRUCTION AND CHARACTERIZATION OF A PILT MUTANT IN
HERBASPIRILLUM SEROPEDICAE SMR1
Vanessa Kessler Chicora, Vânia Carla Silva Pankievicz, Eduardo Balsanelli,
Rose Adele Monteiro, Emanuel Maltempi de Souza, Fábio de Oliveira Pedrosae
Leonardo Magalhães Cruz
Departamento de Bioquímica e Biologia Molecular, Universidade Federal do
Paraná
Herbaspirillum genus is mainly known for its capacity to colonize roots, stems
and leaves of grasses and fix nitrogen. The molecular mechanisms involved in
Herbaspirillum bacteria-plant association are still poorly understood, but
homologous genes to type III secretion system (T3SS), type IV pili (T4P),
adhesins/hemagglutinin and genes involved in lipopolysaccharide (LPS) and
exopolysaccharide (EPS) synthesis were identified in the genome of
Herbaspirillum seropedicae SmR1. Some of these mechanisms have been
elucidated in Herbaspirillum spp. and this work aims to unravel the role of type
IV pili of H. seropedicae motility and plant-bacteria interaction. Previous
transcriptomic analysis confirmed type IV pili genes induction in H. seropedicae
associated with roots of maize and wheat, including pilT gene. The pilT gene,
encoding an ATPase able to retract the pili and develop twitching motility, was
deleted in H. seropedicae. Twenty-nine genes homologous to the pil genes
were identified and are distributed on eight different regions of the genome. The
H. seropedicae PilT protein has four conserved motifs and sequences of the
common amino acids in this protein, however, the C-terminal domain did not
show high degree of conservation. Petri dishes tests with different agar
concentrations was performed to verify how the twitching motility would be
affected. Mutants deleted pilT gene shows growth similar to the wild strain in
1.5% agar, however, in 0.175% agar the mutants showed a greater spread than
wild strain, which can influence the association of bacteria with the plant.
Financial Support: CNPq, INCT
Programa de Pós-Graduação em Ciências (Bioquímica)
39 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ESTROGEN RECEPTOR α GENE (ESR1) EXPRESSION EVALUATION IN
TWO HISTOLOGICAL TYPES OF CANINE MAMMARY TUMOR BY qPCR
Yara de Oliveira Brandão¹, Mariana Busato Toledo¹, Andressa Chequin¹,
Graciele Cristiane Moré Manica¹, Liliane Maria Bacaro Klassen¹, Thierry Grima
de Cristo², Renato Silva de Sousa², Ednéia Amancio de Souza Ramos
Cavalieri¹, Giseli Klassen¹
¹ Laboratório de Epigenética, Departamento de Patologia, Setor de Ciências
Biológicas- Universidade Federal do Paraná
²Laboratório de Patologia Veterinária- Hospital Veterinário- Universidade
Federal do Paraná
Mammary cancer is the mainly neoplasia found in non-spayed bitches Although
the estrogen receptor α (ERα) is one of the molecular marks for breast cancer
in women, it still has a few disagreements about its expression relevance in
canine mammary tumor. In this study the RNA of eight canine mammary
tumors, of which three tubulopapillary carcinoma and five solid carcinoma, was
extracted and used in cDNA synthesis. The transcript quantification was made
by qPCR technique, following the SYBR®Green protocol. The housekeeping
gene was RPS18. Even though solid adenocarcinoma is commonly related to
higher malignance neoplasia degree and many studies have found, by
immunohistochemical technique, lower ERα expression in more aggressive
tumors the qPCR results of this study didn’t show statistical significant
difference in ESR1 expression between both histological types tumors
evaluated. These finds agree with it was previously described in literature
regarding canine mammary adenoma and carcinoma. Once qPCR reaction was
standardized, it is proposed to increase the samples amount and include benign
tumors and non-neoplastic mamma.
Programa de Pós-Graduação em Ciências (Bioquímica)
40 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
CLATHRIN LIGHT CHAIN IN TRYPANOSOMA CRUZI: GENE
IDENTIFICATION AND SUBCELLULAR LOCALIZATION
Ligia Cristina Kalb, Yohana Camila Antunes Frederico, Cassiano Martin Batista,
Iriane Eger, Stênio Perdigão Fragoso and Maurilio José Soares
Laboratory of Cell Biology, Carlos Chagas Institute, Fiocruz-PR
Laboratory of Molecular Biology of Trypanosomatids, Carlos Chagas Institute,
Fiocruz-PR
Department of General Biology, State University of Ponta Grossa
Clathrin-mediated vesicular trafficking, the mechanism by which proteins and
lipids are transported between membrane-bound organelles, accounts for a
large proportion of import from the plasma membrane (endocytosis) and
transport from the trans-Golgi network towards the endosomal system. Clathrin-
mediated events are still poorly understood in the protozoan Trypanosoma
cruzi, the causative agent of Chagas disease in Latin America. In this study,
Clathrin light (TcCLC) chain gene expression and protein localization were
investigated in different developmental forms of T. cruzi (epimastigotes,
trypomastigotes and amastigotes), using polyclonal antibodies raised against T.
cruzi recombinant proteins.
Analysis by confocal microscopy revealed an accumulation of TcCLC at the cell
anterior, where the flagellar pocket and Golgi complex are located. TcCLC
partially colocalized with the Golgi marker TcRAB7-GFP and with ingested
albumin, but did not colocalize with transferrin, a protein mostly ingested via
uncoated vesicles at the cytostome/cytopharynx complex.
In conclusion, Clathrin light chain is expressed in T. cruzi. The protein typically
localize anterior to the kinetoplast, at the flagellar pocket and Golgi complex
region. Our data also indicate that in T. cruzi epimastigotes clathrin-mediated
endocytosis of albumin occurs at the flagellar pocket, while clathrin-independent
endocytosis of transferrin occurs at the cytostome/cytopharynx complex.
Financial Support: CAPES, CNPq, FUNDAÇÃO ARAUCÁRIA, FIOCRUZ
Programa de Pós-Graduação em Ciências (Bioquímica)
41 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ANTICOAGULANT ACTIVITY OF FUCOGALACTAN SULFATED FROM
AGARICUS BISPORUS. OPTIMIZATION OF CHEMICAL SULFATION AND
STRUCTURAL CHARACTERIZATION
Yony Román, Thales R. Cipriani, Marcello Iacomini, Guillerme L. Sassaki
Department of Biochemistry and Molecular Biology, Federal University of
Paraná, Paraná, Brazil
Introduction: Mushrooms have been valued by human kind as an edible and
medical resource containing a number of bioactive molecules with therapeutic
properties, including polysaccharides. Several studies have shown the
anticoagulant activity of sulfated polysaccharides. The common mushroom
Agaricus bisporus is a good source of polysaccharides which can be chemically
sulfated to present anticoagulant activity. Objective: To optimize the chemical
sulfation of a fucogalactan from A. bisporus in order to obtain an anticoagulant
agent.Materials and methods: The fucogalactan was purified from an aqueous
extract of A. bisporus, and characterized by methylation, NMR and HPSEC-
MALLS analyses. It was sulfated in function of different factors [time, molar ratio
of ClSO3H to OH of the polysaccharide (ηClSO3H:ηOH), and weight ratio of
polysaccharide to total reaction volume (Wp/VT)]. The Degree of Sulfation (DS)
was measured and the anticoagulant activity was evaluated by Activated Partial
Thromboplastin Time (APTT) and Prothrombin Time (PT). Discussion and
results: The best anticoagulant activity was obtained with the sulfated
fucogalactan synthesized with a ηClSO3H:ηOH of 18:1 and a Wp/VT of 1:100 in
6 hours of reaction, named E100.. The results of anticoagulant activity of E100
showed a linear increment of APTT for concentrations of 15 to 45 µg.mL-1,
whereas PT was constant between 120 and 160 µg.mL-1. The NMR analyses
suggest that non-reducing end-units of α-L-Fucp and α-D-Galp were greatly
replaced by sulfate groups.Conclusions: E100 inhibited especially the intrinsic
pathway of blood coagulation. Different sulfation conditions generated different
sulfated polysaccharides, which varied at DS and anticoagulant activity.
Financial Support: CAPES, CNPq, Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
42 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
DEEP EUTECTIC SOLVENTS AS NEW MEDIA FOR BIOCATALYSIS:
EXAMPLE OF LIPASE-CATALYZED SYNTHESIS OF PHENOLIPIDS
Erwann Durand
Cirad (La recherche agronomique pour le développement) - Montpellier, França
With the recent interest on green chemistry, the scientists have focused on
developing new and moreefficient solvents to carry out enzymatic-catalyzed
reactions with emphasis on reduced costs, risks and toxicity whileimproving
biodegradability. Among the new available solvents, the multimolecular-based
liquids (such as ionic liquidsand (natural) eutectic solvents) have been the
subject of most recent studies. Currently, and mainly due to its environmental
andeconomic features, (natural) eutectic solvents (NA)DES are arousing much
interest and curiosity. Regarding the biotransformations with lipases, theso-
called “lipophilization” reactions (grafting of lipid moiety onto a hydrophilic
molecule) are of major interest. Indeed, most of the bioactive molecules (e.g.
phenolic acids) express their functionalproperties in a hydrophilic environment,
resulting in a few efficientand advanced applications in formulated-lipid
dispersions.In that sense, lipophilization are particularly advantageousand
effective, and may be seen as a vectorizationkey unlocking the lipid barrier
encountered by the bioactivemolecule while maintaining its original functional
properties.In the case of phenolic acids, the methods commonly used consistin
attaching on the reactive carboxyl group, either a singleor double tail lipophilic
domain (usually an aliphatic witha different carbon backbone) resulting in new
molecule called “phenolipid” withemulsifying properties and often improved
antioxidantactivity.Although these reactions haveshowed tremendous potential,
theyare still complex to implement because of the difficulty in findinga suitable
reactionmedium.Hereby, we propose new perspectivesfor the enzymatic
modification of such polar substrates using the novel generation of green and
inexpensive easy-to-handle (NA)DES.
Programa de Pós-Graduação em Ciências (Bioquímica)
43 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
TARGETINGRESISTANT CANCER CELLS OVEREXPRESSING
MULTIDRUG ABC TRANSPORTERS WITH SELECTIVE DRUG-EFFLUX
INHIBITORS AND APOPTOSIS INDUCERS
Di Pietro, Attilio
BMSSI UMR5086 CNRS-University of Lyon,Institute of Protein Biology and
Chemistry, Passage du Vercors 7, 69367 Lyon, France
MultidrugABC (“ATP-binding cassette”) transporters overexpressed in
chemoresistanttumors are pumping anticancer drugsout of the cells. Totarget
the“breast cancer resistance protein”ABCG2, we have screened different series
of flavonoids and derivatives, such as chalcones,chromones,and
indenoindoles,as inhibitors of mitoxantrone efflux from transfected HEK293
human cells and chemosensitizers of cell proliferation. Two types of selective
and non-competitive inhibitorshave been characterized, either inhibiting or
stimulatingATPase activity.The most potent onewas efficient in vivo on SCID
mice, xenografted with human ABCG2-transfected cells, by chemosensitizing
tumor growth to thedrug-substrateirinotecan. These selective inhibitors
constitute good drug candidates, with low intrinsic toxicity, as sensitizers of cell
proliferation to conventional chemotherapeutics. The “Multidrug Resistance
Protein 1” ABCC1 is able tocatalyze the efflux of glutathione conjugates, or the
co-transport of free glutathione and hydrophobic substrate drugs.Modulators
such as verapamil mimick substratesandinduce a fast and massive efflux of
intracellular glutathione from ABCC1-overexpressing cells,leading to selective
cell death through apoptosis, due to“collateral sensitivity”, or
hypersensitivity.The overexpressed transporter then constitutes the
Achilles’heel of resistant cancer cells. Verapamil being known for cadiotoxic
effects, othermodulatorssuch as xanthones, flavones and flavonoid dimers were
investigated. Glutathione efflux appeared to be necessary, but not sufficient
alone, to trigger apoptosis, indicating the contribution of other partner(s) or
signaling pathway(s). Such apoptosis inducers may constitute a new type of
anticancer drugs operating through an original strategy aimed at selectively
targeting and eliminating multidrug-resistant tumors overexpressing the ABCC1
transporter.
Programa de Pós-Graduação em Ciências (Bioquímica)
44 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
IN SEARCH OF A MODEL SYSTEM TO STUDY ASSOCIATIVE,BIOLOGICAL
NITROGEN FIXATION
Vania C. S. Pankievicz1, Fernanda P. Amaral2, Beverly Agtuca1, Karina
Santos2, Abigail Ferrieri3, Richard Ferrieri4, Maria Berenice R. Steffens1, Ana
Carolina M. Arisi5,Emanuel de Souza1, Fabio Pedrosa1 and Gary Stacey2
1Federal University of Parana, Curitiba, Brazil; 2University of Missouri, Divisions
of Pant Sciences and Biochemistry, C.S. Bond Life Science Building, Columbia,
MO, USA;3Environmental and Molecular Science Laboratory, Pacific Northwest
National Laboratory, Richland, WA, USA;4Biosciences Department, Brookhaven
National Laboratory, Upton, NY 11973, USA; 5University of Santa Catarina -
UFSC, Dept. of Genetic Plant Resources, Florianopolis, Brazil
The world is facing a growing challenge with increasing population,
changing climate and a finite supply of arable land. Hence, perhaps no time in
our history has there been a greater need to invest in agricultural research to
develop sustainable cropping systems to meet the rising demand for food.
Nitrogen is an essential nutrient forplant growth. However, industrial production
of nitrogen fertilizerutilizes fossil fuel with the potential to negatively impact the
environment (e.g., through nitrogen runoff). Legumes and some grasses have
the ability to obtain nitrogen through biological nitrogen fixation (BNF) in
association with soil bacteria. For example, Azospirillumbrasilenseand
Herbaspirillumseropedicaeare examples of diazotrophicbacteria that associate
with several grasses and have been shown to release nitrogen to their host as a
result of BNF. However, it remains controversial whether biological nitrogen
fixation (BNF) contributes significantly to the ability of these bacteria to
promoteplant growth. We believe that this controversy can only be addressed
by more mechanistic studies that would be facilitated by adoption of a useful
plant model system. Our collaboration is focused on analyzing model grass
species for their suitability to support such mechanistic studies. For example,
we recently demonstrated that Setariaviridis, a model C4 grass,isstrongly
colonized by nitrogen fixing bacteria resulting in a significant increase of plant
growth, even under nitrogen limitation. Evidence of nitrogen incorporation by
plant was provided through13NN tracer studies. We were able to follow the
incorporation of 13NN by BNF into plant protein, demonstrating direct, plantuse
of the fixed nitrogen. The use of a genetically tractable, grass model system
should facilitate mechanistic studies of BNF in grass species;hopefully
Programa de Pós-Graduação em Ciências (Bioquímica)
45 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
contributing to the development of technologies to alleviate the need for
industrial nitrogen fertilizers in a moresustainable and environmentally friendly
cropping system.
Programa de Pós-Graduação em Ciências (Bioquímica)
46 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
UPDATE THE RESEARCH IN CARBOHYDRATE CHEMISTRY IN THE
DEPARTMENT OF BIOCHEMISTRY AND MOLECULAR BIOLOGY UFPR
Marcello Iacomini
Department of Biochemistry and Molecular Biology UFPR
Research in Carbohydrate Chemistry in Brazil has few groups. The group
of the Department of Biochemistry and Molecular Biology at the Federal
University of Paraná, probably is the most important group developing this type
of research in our country. Research on polysaccharides started in the 60s in
the last century. Gradually this group has improved the research, precisely
because of the arrival of new researchers have come very important to
strengthen the group and also this kind of research. Precisely because new
lines of research were introduced and with it numerous scientific papers were
published in international journals as well as numerous dissertations and theses
have been produced ever-increasing number and especially in research quality.
All research lines generated scientific production and also with high quality
science. Naturally the number of Master's and PhD students ever increasing
over the decades until the present day in this way, were responsible for the
scientific production of Carbohydrate Chemistry Group. In the last two decades
in addition to the Master's students and PhD to postgraduate absorbed
numerous post docs and today in significant numbers are absorbed by the
groups in the most diverse research areas, bringing a high level of research and
thus the results are observed by the originating publications with the
participation of these new researchers. In this way the results achieved by the
Carbohydrate Chemistry Graduate in Biochemistry and Molecular Biology a
promising future in relation to the quality of the science that has been
developed.
Programa de Pós-Graduação em Ciências (Bioquímica)
47 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ENDOCRINE FUNCTION OF FIBROBLAST GROWTH FACTOR 21 (FGF21)
EXPRESSED IN THE LIVER IN HYPERCALORIC DIET-INDUCED OBESITY
Fabiana Rodrigues Silva Gasparin1, Juliana Moraes Mewes1, Eduardo Hideo
Gilglione1, Clairce Salgueiro Pagadigorria1, Karina Sayuri Utsunomya, Amanda
Tomie Ouchida2, Ingrid C. Gaemers3, Ronald P.J. Oude Elferink3, Jorgete
Constantin1 and Emy Luiza Ishii-Iwamoto1*
1Department of Biochemistry, Laboratory of Liver Metabolism, University of
Maringá, Maringá, Paraná, Brazil
2Laboratory of Bioenergetics, Faculty of Pharmaceutical Sciences, University of
São Paulo, Ribeirão Preto, São Paulo, Brazil.
3Tytgat Institute for Liver and Intestinal Research, Academic Medical Center,
University of Amsterdam, Amsterdam, The Netherlands.
Fibroblast growth factor 21 (FGF21) is an atypical member of the FGF21
superfamily that functions as an endocrine hormone. FGF21 is expressed in
liver, brown (BAT) and white (WAT) adipose tissue and pancreas. The
circulating FGF21 is derived mainly from the liver and coordinates a systemic
response to nutritional stress. In this report, we reported a sex-dependent
difference in hepatic FGF21 expression in response to cafeteria diet-induced
obesity. Swiss mice of both sexes were fed a standard diet or a cafeteria diet
for 14 weeks. Body weight gain, adiposity, liver lipid content, serum levels of
FGF21 and expression of several genes related to lipid and glucose metabolism
and antioxidant defense system were measured. In response to cafeteria diet,
males exhibited greater increases in FGF21 expression in the liver, WAT and
BAT, and a higher serum level of FGF21 compared with females. Metabolic
disturbances and lipotoxicity were found only in females, which exhibited
increased liver fat accumulation (steatosis) and increased cellular oxidative
stress. The analysis of the expression of transcript factors and key enzymes
indicated that the steatosis in females was consequence of metabolic changes
in peripheral tissues. An increased expression of receptor 1 for FGF21 in BAT
was found only in males along with increased expression of uncoupling protein-
1 (UCP1) and fat-specific protein 27 (FSFP). These findings indicated
that BAT is an important target for FGF21 action. These data suggested a
central role of hepatokine FGF21 in protecting the males, but not the females,
against steatosis and lipotoxicity-associated disorders in the cafeteria diet-
induced obesity.
Financial Support: CAPES, CNPq, Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
48 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
REGULATION OF ALTERNATIVE NITROGENASE BIOSYNTHESIS IN
AZOTOBACTERVINELANDII
Ray Dixon
Department of Molecular Microbiology, John Innes Centre, Norwich Research
Park, Norwich, NR4 7UH UK
Azotobactervinelandiiis one of the few diazotrophs that can express three
different types of nitrogenase dependent upon metal availability, namely the
conventional molybdenum enzyme (encoded by nifgenes), the vanadium
nitrogenase (encoded by the vnfgene cluster) and the iron-only nitrogenase
(encoded by the anf gene cluster). In addition to the three specific activators,
NifA, VnfA and AnfA, required for transcriptional activation of the molybdenum,
vanadium and iron-only nitrogenase operons respectively, the A.vinelandii
genome encodes an additional homolog of NifA, designated NifA2 and two
homologs of VnfA, designated as VnfA2 and VnfA3. Each of these activator
homologs has a regulatory GAF domain, which in the case of the VnfA
homologs contains conserved cysteine residues thought to bind an iron-sulphur
cluster. A model to explain the complex hierarchy of metal-dependent gene
regulation mediated by the VnfA homologs will be discussed in this talk.Due to
the complexities of metal-dependent regulation and gene redundancy in A.
vinelandii, it has been difficult to determine the precise genetic requirements for
alternative nitrogen fixation. In this study we have utilized Escherichia coli as a
chassis to build an artificial iron-only (Anf) nitrogenase system comprised of
defined anf and nif genes. Using this system, we demonstrate that the pathway
for biosynthesis of the iron-only co-factor (FeFe-co) is likely to be more simple
than the Mo-dependent (FeMo-co) equivalent. This minimal Anf system has
potential implications for engineering diazotrophy in eukaryotes, particularly in
compartments (e.g. organelles) where molybdenum may be limiting.
Programa de Pós-Graduação em Ciências (Bioquímica)
49 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
THE LONG AND SUGARY ROAD: STRUCTURAL DIVERSITY AND
BIOLOGICAL SIGNIFICANCE OF GLYCOSPHINGOLIPIDS IN PATHOGENIC
AND OPPORTUNISTIC FUNGI
Helio K. Takahashi and Anita H. Straus
Division of Glycoconjugate ImmunochemistryDepartment of Biochemistry
Escola Paulista de MedicinaUniversidade Federal de Sao Paulo
Glycosphingo lipids (GSLs) are ubiquitous membrane components and
have key roles in biological systems, acting as second messengers or
modulators of signal transduction by affecting several cell events, ranging from
cell adhesion, cell growth, cell motility, regulation of apoptosis and cell cycle.
Over the last 20 years our laboratory and other research groups elucidated the
glycan and ceramide structures of more than 20 GSLs from several
pathogenic/opportunistic fungi and parasites, using a combination of nuclear
magnetic resonance, gas chromatography, mass spectrometry, as well as other
chemical, biochemical and immunochemical techniques. Fungal GSLs can be
divided in two major classes: neutral GSLs, e.g., galactosyl- and
glucosylceramide, and acidic GSLs, the glycosylinositol-phosphorylceramides
(GIPCs). Glycan structures in fungal GIPCs exhibited significant structural
diversity and distinct composition when compared to mammalian GSLs, e.g.,
the expression of inositol-mannose and inositol-glucosamine cores and the
terminal residue of β-D-galactofuranose which are absent in mammalian cells.
Studies performed by our group demonstrated that GIPC
(Galfβ6[Manα3]Manα2InsPCer) elicited in patients with paracoccidioidomycosis
an immune response with production of antibodies directed to the terminal
residue of β-D-galactofuranose. Further studies also showed that inhibition of
glucosylceramide (GlcCer) biosynthetic pathways affects fungal colony
formation, spore germination and hyphal growth, indicating that enzymes
involved in GlcCer biosynthesis may represent promising targets for the therapy
of fungal infections. Recently, it was shown that GlcCer and GIPCs are
preferentially localized in membrane microdomains and monoclonal antibodies
directed to these GSLs interfere in several fungal biological processes such as
growth and morphological transition. An in-depth knowledge of fungal
microdomain interactions in combination with the elucidation of the concerted
action of membrane microdomain, GSL and cell wall will certainly help to
Programa de Pós-Graduação em Ciências (Bioquímica)
50 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
elaborate a more refined concept of key events related to survival and
proliferation of parasite and pathogenic/opportunistic fungi in the human host.
Programa de Pós-Graduação em Ciências (Bioquímica)
51 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ANTI-TUMOR COMPOUNDS: DIFFERENT APPROACHES TO INVESTIGATE
THE MECHANISMS OF ACTION
Maria Eliane Merlin Rocha, Silvia Maria Suter Correa Cadena, Glaucia Regina
Martinez, Guilhermina Rodrigues Noleto, andSheila Maria Brochado
Winnischofer
Department of Biochemistry and Molecular Molecular Biology, UFPR
Cancer treatments include surgery radiation, chemotherapy, hormone
therapy, immune therapy, and targeted therapy. Furthermore, the existing
pharmacological-based treatments are insufficiently effective and generate
many side effects. It is necessary new drugs to treatment of cancer. In the
Center of Studies in Bioenergetics and Biochemistry of Drugs and Xenobiotics
we study molecular mechanisms of action of flavonoids, mesoionic compounds,
statins and other xenobiotics in tumor various cell lines. New drugs studies of
structure-activity relationship and pro-oxidants on tumor cells effects are
investigated. Involvement of mitochondrial pathway on antitumor derivatives
1,3,4 thiadiazoles mesoionic; polysaccharides effects on modulation of immune
system are other focus of studies. Photodynamic action using dyes and UV
radiation to verify DNA damage promoved by singlet oxygen in melanoma cells
and analyses of role of the RECK tumor suppressor gene and their alternative
transcripts are verified too. Compounds studied can induce different cell
responses as cell death; increase of ROS levels; senescence; autophagy;
alteration of cell cycle progression; inhibition of cellular migration or are
imunomodulators. Furthermore, we have been studying the inhibition produced
by flavonoids on drug efflux by ABCG2.
Financial Support: CNPq, CAPES, INCT-REDOXOMA and Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
52 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
FORMATION OF PROTEIN PARTICLES AND THEIR INFLUENCE ON THE
STRUCTURE AND RHEOLOGY OF POLYSACCHARIDE SOLUTIONS AND
GELS
Bach T. Nguyen, Christophe Chassenieux, Lazhar Benyahia, Taco Nicolai
LUNAM, Université du Maine, IMMM UMR CNRS 6283, PCI, 72085 Le Mans
cedex 9, France
β-lactoglobulin (β-lg) is a globular protein and is the major protein
component of whey. β-lg aggregates irreversibly when heated and gels above a
critical concentration. In a narrow range of conditions heated β-lg forms
spherical microgels the size of which can be controlled by the pH and the
concentrations of added CaCl2. Here I will discuss how the presence of protein
microgels influences the behaviour of the polysaccharide -carrageenan (κ-car).
κ-car gels reversibly below a critical temperature that can be reduced by adding
small amounts of specific ions such as K+. κ-car is sometimes added to milk
based products in order to improve the texture. The aggregation and gelation of
both κ-car and β-lg are very sensitive to the presence of calcium ions that are
most often present in the food products. Therefore it is important to understand
the effect of Ca2+ on gelation of mixtures. The effect of adding CaCl2 to mixtures
of κ-car and β-lg microgels at neutral pH on the morphology and the elasticity is
presented. The β-lg microgels were formed by heating either before or after
mixing with κ-car. It will be shown that both β-lg and κ-car specifically bind Ca2+
and that in mixtures they compete for Ca2+. This influences aggregation and
gelling of each biopolymer. In addition, thermodynamic incompatibility drives
micro phase separation above a critical κ-car concentration with β-lg microgels.
An attempt is made to disentangle the effects of competition for Ca2+ and
thermodynamic incompatibility on the structure and the elasticity of the
mixtures.
Programa de Pós-Graduação em Ciências (Bioquímica)
53 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
GLYCOCONJUGATES FROM PATHOGENIC FUNGI
Mariana I. D. S. Xisto1, Vera C. B. Bittencourt2, Livia C. Liporagi-Lopes3, Rosa
M. T. Haido2, MorenaS. A. Mendonça 4, Guilherme Sassaki5, Rodrigo T.
Figueiredo6 , Maria Teresa V. Romanos 7 and Eliana Barreto-Bergter1
1Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de
Góes – UFRJ, Rio de Janeiro, Rio de Janeiro, Brazil
2Departamento de Microbiologia e Parasitologia, Instituto Biomédico, UNIRIO
3Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia -
UFRJ, Rio de Janeiro, Rio de Janeiro, Brazil
4Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, RJ
5Departamento de Bioquímica e Biologia Molecular, Universidade Federal do
Paraná, Curitiba, Paraná.
6Campus de Xerém, Universidade Federal do Rio de Janeiro, Instituto de
Ciências Biomédicas, Universidade Federal do Rio de Janeiro
7Departamento de Virologia, Instituto de Microbiologia Paulo de Góes – UFRJ,
Rio de Janeiro, Rio de Janeiro, Brazil
A peptidorhamnomannan (PRM) that consists of a peptide chain
substituted both by O- and N-linked glycans was isolated from the cell wall of S.
prolificans, an emerging opportunistic fungus, with remarkable resistance to
antifungal agents and able to cause localized infection in immune competent
patients and disseminated infection with high rate of mortality among immune
compromised patients. Its chemical structure was elucidated using methylation
analysis and 13C - nuclear magnetic resonance. The importanceofO-
linkedoligosaccharidespresent in peptidorhamnomannan (PRM)
fromthecellwallof the fungusScedosporiumprolificans for
recognitionandphagocytosisofconidiabymacrophageswasanalyzed.Adding PRM
ledto a dose-dependentinhibitionofconidiaphagocytosis, whereas de-O-
glycosylated PRM didnot show anyeffect. PRM inducedthe release
ofmacrophage-derivedantimicrobialcompounds. However, O-
linkedoligosaccharides do notappeartoberequired for such induction. The
effectof PRM onconidia-
inducedmacrophagekillingwasexaminedusinglatexbeadscoatedwith PRM or de-
Programa de Pós-Graduação em Ciências (Bioquímica)
54 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
O-glycosylated PRM. A decrease in macrophageviability similar
tothatcausedbyconidiawasdetected. However,
macrophagekillingwasunaffectedwhenbeadscoatedwith de-O-glycosylated PRM
wereused, indicatingthetoxiceffectofO-linkedoligosaccharidesonmacrophages.
In addition, PRM triggered TNF release bymacrophages. ChemicalremovalofO-
linkedoligosaccharidesfrom PRM abolishedcytokineinduction,
suggestingthattheO-linkedoligosaccharidicchains are importantmoietiesinvolved
in inflammatory responses throughtheinductionof TNF-α secretion. In summary,
we show thatO-glycosylation plays a role in therecognitionanduptakeofS.
prolificansbymacrophages, killingofmacrophagesandproductionofpro-
inflammatorycytokines.
Financial Support: CNPq, CAPES, FAPERJ, UFRJ
Programa de Pós-Graduação em Ciências (Bioquímica)
55 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
SEARCHING FOR TARGET MOLECULES FOR DIAGNOSTICS AND
THERAPEUTICS OF DEGENERATIVE DISEASES (CANCER, DIABETES)
AND REGENERATIVE MEDICINE
Marina Trombetta-Lima, Fernando Henrique Lojudice, Aline Maia Lobba, Ana
Claudia Oliveira Carreira, Sheila Maria Brochado Winnischofer, Renato Astorino
Filho, Túlio Felipe Pereira, Patricia Mayumi Kossugue, Fernando Janczur
Velloso, Raquel Arminda Carvalho Machado, Marluce da Cunha Mantovani,
Suely Kazue Marie, Maria Lucia Correa Giannella, Mari Cleide Sogayar
NUCEL/NETCEM Cell and Molecular Therapy Center, Internal Medicine Dept.,
Medical School, University of São Paulo, São Paulo, 055360-130 SP, Brazil
The incidence of degenerative diseases, such as cancer and diabetes
mellitus, has been increasing at alarming rates due, mainly, to greater life
expectancy, population ageing and obesity. Therefore, tools for early diagnosis
and efficient therapeutics are essential.We focused on the deadliest tumorof the
central nervous system, namely: Glioblastoma (GBM), using both a rat glioma
cellular model and human tumor samples to search for genes involved in the
action of chemotherapeutics (glucocorticoids-GCs) and in chemoresistance to
these drugs, and, also, on the role played by the RECK tumor and metastasis
suppressor gene alternatively spliced variants, finding important correlations
with patients survival rates and prognosis.Straighforward, uncontroversial,
molecular markers for mammary carcinoma are still lacking, but the recent
tumor stem cell theory provided some insights that led us to find two different
stem cell markers with high diagnostic and therapeutic potential for these
tumors.Insulin-dependent or type 1 diabetes mellitus arises from the auto-
immune destruction of insulin-producing pancreatic beta-cells. Exogenous
insulin replacement does not perfectly mimick the physiological blood glycemic
control, leading to renal failure, cardio-vascular diseases, blindness and other
problems. Whole pancreas and pancreatic islet transplantation are possible
therapeutic options, however, both require immunossupression of the patients.
Islet encapsulation, stem cell differentiation into insulin producing cells and
Tissue Bioengineering are promising new alternatives arising from
Regenerative Medicine, which is based on: a) isolated and well-characterized
stem cells; b) recombinant peptide growth factors; c) extracellular matrix
scaffolds. Personalized therapeutics, tissue repair/reconstruction and organs
Programa de Pós-Graduação em Ciências (Bioquímica)
56 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
replacement, employing different cell types, growth factors and 3D printers are
concrete perspectives for the near future.
Financial Support: BNDES, CNPq, CAPES, FAPESP, MCTI, MS-DECIT
Programa de Pós-Graduação em Ciências (Bioquímica)
57 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
BIOLOGICAL NITROGEN FIXATION- A FRUITFUL IDEA
José Ivo Baldani
Embrapa Agrobiologia, Km 07, 23891-000 – Seropédica, RJ
Biological nitrogen fixation (BNF) is a natural process realized by a small
group of Prokaryotes that lives in terrestrial and aquatic environment. Their
contribution to the global nitrogen cycle vary from 100 to 300 million tons N/year
with about 1/3 derived from terrestrial microorganisms and the rest from the
oceans. The main sources of terrestrial contribution are from the association of
rhizobia with legumes and of associative/endophytic bacteria with
graminaceous plants. Soybean is the best example of the symbiosis mainly in
Brazil where all the nitrogen required for high yield comes from the BNF and
provides an economy of about U$ 10 billion with N fertilizer. Other legumes
such as common beans and cowpea are also benefited from the symbiosis with
rhizobia including the new species such as Microvirga vignae described
recently. In addition, the recovery of mine areas using the tripartite symbiosis
(legume tree, rhizobia and micorrhizae) reinforce the importance of the
biological nitrogen fixation to the environment. On the other hand, the BNF
contribution of the associative/endophytic association is lower; nevertheless, the
economy with N fertilizer application is quite substantial such it was
demonstrated for sugarcane. In contrast, the BNF contribution to cereals (rice,
sorghum, wheat and corn) does not meet the N requirement despite the
diversity of nitrogen–fixing bacterial species (Gluconactobacter diazotrophicus,
Azospirillum lipofereum, A. brasilense, Nitrospirillum amazonense,
Herbaspirillum seropedicae and Burkholderia tropica ) that have been isolated
and tested in association with these cereals. Therefore, strains more efficient
and competitive should be selected including other additional characteristics (N-
use efficiency, phytohormones and siderophere production, Pi and Zn
solubilization, etc). About 3,000 strains, phenotypically characterized as rhizobia
and associative/endophytic bacteria, are deposited at the Embrapa Agrobiology
Culture Collection and now are molecularly and physiologically characterized
envisaging the creation of the Genetic Resource Center-CRB Johanna
Döbereiner at Embrapa and their bioprospection for agriculture application.
Financial Support: CNPq, CAPES, FAPERJ and Embrapa.
Programa de Pós-Graduação em Ciências (Bioquímica)
58 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
FROM STRUCTURE TO CATALYSIS: RECENT ADVANCES IN THE
BIOTECHNOLOGICAL APPLICATIONS OF LIPASES
Erika C. G. Aguieiras and Denise M. G. Freire
Lipases are highly appreciated as biocatalysts due to their peculiar
characteristics such as the ability to utilize a wide range of substrates, high
activity and stability in organic solvents, and regio- and/or enantioselectivity.
These biocatalysts account for 5% of the global market for industrial enzymes
and can be applied in a variety of biotechnological processes, including food
modification, detergent formulation, cosmetics, pharmaceutical, leather, textile,
and paper industries, biodiesel and biopolymer synthesis, or pretreatment of
lipid-rich wastewaters. However, in certain segments of industry such as
biodiesel production and treatment of wastewaters, the use of lipases is still
limited by their high cost. Thus, there is a great interest in obtaining low- cost,
highly active, and stable lipases that can be applied in several different
industrial branches. The use of low-cost enzyme preparations obtained by solid
state fermentation (SSF) and application of the solid enzymatic preparation
(SEP) is an alternative for reduction of costs with extraction, purification and
enzyme immobilization. In addition, by SSF is possible to remove toxic and/or
anti-nutritional compounds and to enhance the nutritional value of the fermented
solids through protein enrichment of the residues, that can be potential
substitutes for animal feed components. Moreover, currently the design of
specific enzymes for each type of process has been used as an important tool
to address the limitations of natural enzymes. Nowadays, with the progress in
protein engineering and structure-based rational design it is possible to “order”
a “customized” lipase that has ideal properties for the development of the
desired bioprocess.
Programa de Pós-Graduação em Ciências (Bioquímica)
59 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
AN OMICS PERSPECTIVE OF BENEFICIAL CEREAL-DIAZOTROPHIC
BACTERIA INTERACTIONS
Emanuel Maltempi de Souza
Núcleo de Fixação de Nitrogênio
Department of Biochemistry and Molecular Biology, UFPR.
Nitrogen is main factor limiting plant growth and productivity. However,
increasing amounts of nitrogen fertilizers cannot be indefinitely used to meet the
demandfor world food production due to high costs and environmental impacts.
Two major groups of diazotrophic bacteria associate closely with plants and can
transfer fixed nitrogen. The symbiotic organisms that form differentiated
structures on plant roots and the associative bacteria that colonize roots both
epiphytically and endophytically. Both groups can promote plant growth and
improve productivity in a sustainable way. The mechanism through which
symbiotic association is formed and the exchange of nitrogen and carbon
between plant and bacteria is well studied, and this group of bacteria is largely
used in agriculture. On the other hand, the molecular mechanisms of plant
recognition, colonization and nutrient exchange between diazotrophic
epiphytes/endophytes and plants are hardly known. Azospirillumbrasilenseand
Herbaspirillumseropedicae are promising plant growth promoting bacteria
capable of colonizing roots of important cereals. We used RNA-Seq
transcriptional profiling and proteomics to further the understanding of the
association of wheat, maize and rice with these bacteria. Genes
codingplantdefence proteins, nutrient uptake andsecondary metabolites were
differentially expressed in response to the bacteriam while the bacteria express
nitrogenfixation,cellmotilityandcellwallbiogenesis genes. These studies
represent an advancement in the understanding of the diazotrophic bacteria-
cereals interaction.
Financial Support: CNPQ/INCT-FBN, CAPES and Fundação Araucária
Programa de Pós-Graduação em Ciências (Bioquímica)
60 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
NEW LIPASES FROM METAGENOMIC LIBRARIES: FROM
CHARACTERIZATION TO THE DEVELOPMENT OF APPLICATIONS
Nadia Krieger
Department of Biochemistry and Molecular Biology, UFPR
In recent years, the application of traditional methods for the isolation and cultivation of microorganisms in samples collected from the environment has resulted in a high rate or rediscovery of known enzymes that have already been well characterized. These enzymes do not necessarily have characteristics suitable for use in reaction media containing organic solvents that are often used in biocatalysis. On the other hand, since metagenomic prospection allows access to bacteria that are not cultivatable using traditional techniques, it has a good chance of identifying new enzymes. The metagenomic technique consists of the extraction of DNA fragments from samples collected from the environment, with the construction of a library of clones, called a “metagenomic library”. The cloned genes are sequenced and, in the case of enzymes, screened for activity and the ability to maintain stability under the proposed reaction conditions. This talk will show how we have used the metagenomic technique to isolate new lipases with interesting characteristics that may allow their use in the organic reaction media and, consequently, in biocatalytic processes.
Programa de Pós-Graduação em Ciências (Bioquímica)
61 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ÍNDICE DE AUTORES
Abreu, E.C.A. 13
Agtuca, B. 44
Aguieras, E.C.G. 58
Amaral, F.P. 44
Aranha, E.M. 14; 19
Araújo, P.H.H. 34
Arisi, A.C.M. 44
Baldani, J.I. 57
Balsanelli, E. 11; 38
Barbieri, S F. 37
Barddal, H.P.O. 23
Bark, J.M. 10
Barreto-Bergter, E. 53
Batista, C.M.40
Batista, L.A.C. 17
Batista, M.B. 28
Benyhmia, L. 52
Bezerra, I.L. 6; 25
Biembengut, I.V 22; 26
Bittencourt, V.C.B. 53
Bonato, P. 31
Brandão, Y. O. 8; 22; 26; 39
Brandt, A.P. 9
Brown, G.G. 16
Cadena, S.M.S.C. 9; 18; 24; 32; 51
Caillot, A. R. C.25
Camilios-Neto, D. 31
Canuto, V.C.9
Carreira, A.C.O.
Carvalho, M.M. 19; 30
Cassilha, B.A.R. 29
Cavalieri, E.A.R.39
Chandra, G. 28
Chassenieux, C. 52
Chaves, P.F.P.36
Chequin, A. 22; 26; 39
Chicora, V.K. 38;
Chubatsu, L.S. 16; 31
Cipriani, T.R. 7; 23; 41
Colodi, F.G. 19; 30
Constantin, A. 47
Cordeiro L.M.C. 7; 36
Cordeiro, E.W.F. 17
Cristo, T.G.C.39
Cruz, L.M. 38
Cruz, O.M. 10
Programa de Pós-Graduação em Ciências (Bioquímica)
62 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
De Godoy, R. C. B. 37
Di Mascio, P. 32
Di Pietro, A. 43
Dias, G.S. 21
Dixon, R. 28; 48
Ducatti, D.R.B. 14; 15; 19; 20
Durand, E. 42
Echevarria, A.9
Eger, I. 40
Elferink, R.P.J.O. 47
Esteves, E.D.16
Faoro, H.29
Fernandes, K.L.M. 24
Ferrieri, A. 44
Ferrieri, R. 44
Feuser, P.E. 34
Figueiredo, R.T. 53
Filho, R.A.
Floh, E.I.S. 18
Fortes, F. 18
Fragoso, E.P. 40
Franco, C.R.C. 27
Frederico, Y.C.A. 40
Freire, D.M. 58
Freitas, R.A. 30
Furlanetto, A.L.D.M. 18
Gaemers, I.C. 47
Gasparin, F.R.S. 47
Gianella, M.L.C. 55
Gilglione, E.H. 47
Gonçalves, A.G. 14; 15; 20
Gracher, A.H.P. 23
Haido, R.M.T. 53
Heinrich,T.A. 35
Heuke, J.G. 14
Hilgert, R.M. 17
Huergo, L.F. 29
Iacomini, M.7; 23; 36; 41; 46
Ishii-Iwamoto, E.L 47
Kalb, L.C. 40
Kaziuk, F.D.18
Klassen, G. 8; 22; 26; 39
Klassen, L. M. B. 8; 22; 26; 39
Kossugue, P.M. 55
Kozlowski, E.O. 12
Krieger, N. 21; 60
Lammel, D.R. 11
Lana, P.C. 11
Liporagi-Lopes, L.C. 53
Lobba, A.M. 55
Programa de Pós-Graduação em Ciências (Bioquímica)
63 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
Lojudice, F.H. 55
Lucinda-Silva, R.M. 27
Luz Jr., L.F.L. 21
Machado, A.C. 55
Manica, G. C. M. 8; 22; 26; 39
Mantovani, M.C. 55
Marie, S.K. 55
Marinez, A. 12
Martinez, G.R. 10; 32; 35; 51
Mazepa, E. 15
Mendonça, M.S.A. 53
Mewes, J.M. 47
Mitchell, D.A. 21
Monteiro R.A. 28; 38
Nassato, P.L. 33
Nguyen, B.T 52
Nicolai, T. 52
Noleto, G.R. 13; 24; 51
Noseda, M.D. 14; 15; 19; 20; 30; 33
Noseda, M.E.R.D. 14; 15; 19, 30; 33
Oliveira A.F. 7
Oliveira, M.A.S. 22
Ouchida, A.T. 47
Pagadigorria, C.S. 47
Paim, C.S. 17
Pankievicz, V. 38; 44
Pavao, M.S.G. 12
Pedrosa, F.O. 11; 16; 29; 31; 38; 44
Pereira, T.F. 55
Peres, P.S. 32
Petkowicz, C.L.O. 24; 27; 37
Pignon, F. 33
Pino-Gomes, R. 10; 15
Pires, A. R. A. 9
Ramos, E. A. S. 8; 22; 26; 39
Restrepo, D.C. 12
Ribeiro, C.S.P. 10
Ricci-Júnior, E. 34
Rigo, L. 31
Rinaudo, M. 33
Rios, J.C. 12
Rocha, M.E.M. 51
Román, Y. 41;
Romanos, M.T.V. 53
Ruthes, A.C. 37
Santana-Filho, A. P. 6; 25
Santos, A.L.W. 18
Santos, G.C.S. 20
Santos, K. 44
Sassaki, G. L. 6; 25; 41; 53
Programa de Pós-Graduação em Ciências (Bioquímica)
64 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
Sayer, C. 34
Scalfo, A.C. 32
Scarduelli, M. 29
Seccon, D.M. 11
Silveira, J.L.M. 33; 37
Simas-Tosin, F.F. 23
Soares, M.J. 40
Sogayar, M.C. 55
Sousa, R.S.39
Souza, E. M. 8; 11; 16; 22; 26; 28; 29;
31; 38; 44; 59
Stacey, G. 44
Stefens, M.B.R. 44
Stopiglia, C.D.O. 17
Straus, A.M. 49
Szwarc, P. 35
Tadra-Sfeir, M. Z. 11; 31
Tagahashi, M.K 49
Teixeira, F.C.O.B. 12
Toledo, M. B. 8; 22; 26; 39
Trombetta-Lima, M.
Utsunomya, K.S. 47
Valerio, A. 32
Velloso, F.J. 55
Vriesmann, L.C. 27
Wassem, R. 11; 31
Winnischofer, H. 10
Winnischofer, S.M.B. 10; 15; 32; 51; 55
Xisto, M. 53
Programa de Pós-Graduação em Ciências (Bioquímica)
65 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
ANEXO
PROGRAMAÇÃO DO EVENTO
Quarta-feira – 23 de setembro de 2015
18:00 Abertura e Coquetel
Local: Teatro da Reitoria - UFPR
Rua XV de Novembro, 1299 – Centro - Curitiba – PR
***
LOCAL dos Simpósios e cerimônia de encerramento: Auditório do Setor
de Ciências Sociais Aplicadas – Campus do Jardim Botânico
Av. Pref Lothario Meissner, 3400 – Jardim Botânico
***
Quinta-feira - 24 de setembro 2015
9:00 – 12:00 Simpósio Biotecnologia
Coordenador: David Mitchell
Departamento de Bioquímica e Biologia Molecular – UFPR
Programa de Pós-Graduação em Ciências (Bioquímica)
66 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
“Deep eutectic solvents as new media for biocatalysis: Example of lipase-
catalyzed synthesis of phenolipids”
Palestrante : Erwann Durand
Cirad (La recherche agronomique pour le développement) - Montpellier, França
“From structure to catalysis: recent advances on the biotechnological
applications of lipases”
Palestrante :Erika Cristina G. Aguieiras
UFRJ (Instituto de Química) - Rio de Janeiro
“New lipases isolated from metagenomic libraries: from characterization to the
development of applications”
Palestrante :Nadia Krieger
Departamento de Química – UFPR
Quinta-feira - 24 de setembro 2015
12:00 – 14:00 Sessão de Pôsteres
14:00 – 17:00 Simpósio Fixação Biológica de Nitrogênio
Coordenador: Fábio Oliveira Pedrosa
Departamento de Bioquímica e Biologia Molecular – UFPR
“Biological Nitrogen Fixation, from the discovery of nif gene to nitrogen fixation
in planta”
Palestrante :Ray Dixon
John Innes Centre – Inglaterra
Programa de Pós-Graduação em Ciências (Bioquímica)
67 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
“Nitrogen fixation in non-leguminous plants”
Palestrante :Gary Stacey
Missouri University – Estados Unidos
“Fixação biológica de nitrogênio: uma ideia fértil”
Palestrante :Ivo Baldani
Embrapa –Rio de Janeiro Agrobiologia
“Fixação de nitrogênio: uma perspectiva ômica”
Palestrante :Emanuel Maltempi de Souza
Departamento de Bioquímica e Biologia Molecular – UFPR
Sexta-feira - 25 de setembro 2015
9:00 – 12:00 Simpósio Bioquímica Farmacológica
Coordenador: Maria Eliane Merlin Rocha
Departamento de Bioquímica e Biologia Molecular – UFPR
“Targeting resistant cancer cells overexpressing multidrug ABC transporters
with selective drug-efflux inhibitors and apoptosis inducers”
Palestrante :Dr. Attilio Di Pietro
Institut de Biologie et Chimie des Protéines (IBCP)- Université Lyon 1 (França)
Programa de Pós-Graduação em Ciências (Bioquímica)
68 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
“Busca de moléculas-alvo para doenças degenerativas (cancer, diabetes) e
Medicina Regenerativa”
Palestrante : Mari Cleide Sogayar
Universidade de São Paulo
“Função endócrina do fator de crescimento de fibroblastos 21 (FGF21)
expresso no fígado na obesidade induzida por dieta hipercalórica”
Palestrante : Emy Luiza Ishii Iwamoto
Universidade Estadual de Maringá
“Anti-tumorais: diferentes abordagens na investigação dos mecanismos de
ação”
Palestrante : Maria Eliane Merlin Rocha
Departamento de Bioquímica e Biologia Molecular – UFPR
Sexta-feira - 25 de setembro 2015
12:00 – 14:00 Sessão de Pôsteres
14:00 – 17:00 Simpósio Estrutura e Propriedades de Carboidratos
Coordenador: Marcello Iacomini
Departamento de Bioquímica e Biologia Molecular – UFPR
“Formation of protein particles and their influence on the structure and rheology
of polysaccharide solutions and gels”
Palestrante :Taeke Nicolai
Institute des Molecules et des Materiaux – CNRS Maine – France
Programa de Pós-Graduação em Ciências (Bioquímica)
69 LIVRO DE RESUMOS DO SIMPÓSIO COMEMORATIVO DOS 50 ANOS DO PROGRAMA DE
PÓS-GRADUAÇÃO EM CIÊNCIAS (BIOQUÍMICA) UFPR _ setembro/2015
“Glicoconjugados de fungos patogênicos”
Palestrante : Eliana Barreto Bergter
Instituto de Microbiologia Paulo de Góes, UFRJ
“The long and sugary road: Structural diversity and biological significance of
glycosphingolipids in pathogenic and opportunistic fungi”
Palestrante:Helio K. Takahashi
Escola Paulista de Medicina - Universidade Federal de Sao Paulo
“Atualização da Pesquisa em Química de Carboidratos no Departamento de
Bioquímica e Biologia Molecular”
Palestrante: Marcello Iacomini
Departamento de Bioquímica e Biologia Molecular – UFPR
Sexta-feira - 25 de setembro 2015
17:30 Cerimônia de encerramento