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Abstracts / Journal of Biotechnology 185S (2014) S37S125

acid (C22:63, DHA), play multifunctional roles in human health

and diseases. The traditional source of these fatty acids is sh.
Since sh stocks are in global decline, new sources of omega-3
LC-PUFA have to be found. The aim of the present study was to
nd potential alternative sources of these fatty acids in the territory of The Republic of Kazakhstan. Twelve strains of lamentous
fungi isolated from soil and water were screened for lipids and LCPUFA: Penicillium sp., Penicillium restrictum, Penicillium aculeatum,
Penicillium anatolicum, Penicillium bilaiae, Penicillium commune,
Penicillium raistrikii, Penicillium crustosum, Fusarium sp. (2 strains),
Aspergillus sp. and Mucor circinelloides. These fungi were grown
in Sabouraud medium for 10 days. Lipid contents of these strains
ranged from 1.5% to 7.4% of dry weight. No LC-PUFA were found in
P. commune, P. crustosum, Aspergillus sp., M. circinelloides, whereas
other strains contained EPA ranging from 1.0% to 3.5% of the total
fatty acids, P. restrictum accumulated the largest amount. The cultural conditions that can increase the production of EPA in P.
restrictum (e.g. the addition of glucose, glycerin and zinc) are currently under investigation.
http://dx.doi.org/10.1016/j.jbiotec.2014.07.332
Circulating tumor cells (CTCs) in association to
breast cancer subtypes and inammatory
pattern
Angela Toss 1, , Jocelyn A. Sendecki 2 , Tiffany P.
Avery 2 , James M. Reuben 3 , Massimo
Cristofanilli 2
1 University of Modena and Reggio Emilia, Modena,
Italy
2 Thomas Jefferson University, Philadelphia, PA, USA
3 The University of Texas MD Anderson Cancer
Center, Huston, TX, USA

The phytochemical, antibacterial and

antioxidant activity of wild plants growing in
Kazakhstan against diabetic foot ulcer
Tatyana Kustova 1, , Tatyana Karpenyuk 1 , Alla
Goncharova 1 , Samir Ross 2
1 Faculty of Biology and Biotechnology, Al-Farabi
Kazakh National University, Almaty, Kazakhstan
2 School of Pharmacy, NCNPR, The University of
Mississippi, University, USA

E-mail address: kus talya@yahoo.com (T. Kustova).

Diabetic foot infection (DFI) is one of the most frequent and
complex problems among patients suffering from diabetes. DFI
is polymicrobial in nature. Moreover most of the bacteria have
multidrug resistance. Therefore, much attention has been paid to
nd biological active substances from plants that help to solve
this problem. Thus, the aim of this study was to evaluate phytochemical, antibacterial and antioxidant activity of crude extracts
isolated from plants growing in Kazakhstan: Epilobium hirsutum
L. (Onagraceae), Rumex confertus Wiild. (Polygonaceae), Vexibia
alopecuroides (L.) Jakovl. (Fabaceae). The results of the present
research clearly indicate that the antibacterial and antioxidant
activities vary with the plant species. Among the different crude
extract that were evaluated, the dichloromethane extract of roots
of V. alopecuroides showed the strongest activity against Staphylococcus aureus (IC50 3.05 g/mL) and methicillin-resistant S. aureus
(IC50 2.9 g/mL). Despite the fact that the extract of V. alopecuroides
has not shown high antioxidant activity, it is still highly promising
for the study because it showed high antimicrobial activity. Highest antioxidant activity (<85% inhibition of DPPH) was shown by
E. hirsutum L. and R. confertus Wiild. Based on the research results
plants were selected to produce drugs from their extracts.
http://dx.doi.org/10.1016/j.jbiotec.2014.07.334

E-mail address: angela.toss@tin.it (A. Toss).

Circulating tumor cells (CTCs) are an independent prognostic factor
for progression-free (PFS) and overall survival (OS) in metastatic
breast cancer (MBC). The present analysis evaluates the prognostic value of CTCs in HER-2 positive disease receiving targeted
therapies. This is a retrospective analysis of 516 patients with
MBC from the MDACC database. The PFS and OS were evaluated
in association to baseline CTCs (<5 vs. 5) within the different subtypes (HR+/HER2, HR+/HER2+, HR/HER2+, TNBC) and
inammatory breast cancer (IBC). Among 516 patients, 56% had
HR+/HER2 tumor, 10% HR+/HER2+, 10% HR/HER2+, and 24%
were TNBC, including 25% with IBC. Overall, patients with CTCs 5
had a signicantly lower median survival time (HR = 2.00). In
TNBC, HR+/HER2 and HR+/HER2+ tumors, CTC 5 were signicantly associated with lower survival times (HR = 1.96, 2.56 and
3.15 respectively). HR/HER2+ subtype and IBC demonstrated no
differences in survival according to baseline CTCs enumeration.
Baseline enumeration have strong prognostic value except for
HR/HER2+ subtype treated with targeted therapy and IBC, suggesting that CTCs are more homogeneous in expression of HER2
before treatment, while patients with resistant disease may have
cells with epithelialmesenchymal transition that are undetectable
by CellSearch , suggesting the need to introduce new detection
methods.
http://dx.doi.org/10.1016/j.jbiotec.2014.07.333

Molecular cloning, over-expression,

purication, and functional characterization of
the meningococcal HrpA protein C-terminal
domain
Graziano Pizzolante , Roberta Romano, Carla
Palumbo, Pietro Alifano
Dipartimento di Scienze e Tecnologie Biologiche ed
Ambientali, Universit del Salento, Lecce, Italy
E-mail address: graziano.pizzolante@unisalento.it (G. Pizzolante).
The two-partner secretion (TPS) systems of Gram-negative bacteria consist of a large secreted exoprotein (TpsA) and a transporter
protein (TpsB) located in the outer membrane. The functions
identied for the TpsA proteins are numerous and contribute to
bacterial tness or virulence. Three different TPS systems in Neisseria meningitidis (meningococcus) are present. A previous study
demonstrated that the meningococcal HrpBHrpA TPS system was
essential for intracellular growth/survival and escape from infected
cells. Interestingly, the hrpA genes display signicant C-terminal
sequence variation. In this study, we highlighted by bioinformatics
tools the presence of a putative hemolysin/hemagglutinin at the Cterminal domain of the HrpA protein. We cloned, over-expressed,
puried and tested the protein activity on the human erythrocytes.
The hemolytic activity assay showed that the C-terminal moiety
induced erythrocytes lysis with MnCl2 . Instead, no hemolytic activity was recorded either in absence of ion supplement or in the
presence of magnesium. Our data suggest that the HrpA protein Cterminal domain is a manganese-dependent lysin directly used by

Abstracts / Journal of Biotechnology 185S (2014) S37S125

meningococci to escape from vacuoles and invade neighbors cells.

Thus, understanding the molecular mechanisms of the meningococcal TPS system may help discover when and how a transitory
colonizer of the nasopharynx as N. meningitidis became human
pathogen.

The inuence of resveratrol on hemoglobin

system and change in erythrocytes structure
under oxidative processes induction
Marina Yurievna Yazykova , Viktor Vasilevich
Revin, Kseniya Andreevna Borisova

http://dx.doi.org/10.1016/j.jbiotec.2014.07.335

Department of Biotechnology, Samara State

University, Samara, Russia

Anti-quorum sensing activities of different

extracts from Chlorella vulgaris

E-mail address: yazykovasamgu@mail.ru (M.Y. Yazykova).

Kbra Erkan 1, , Demet Erdnmez 1 , Minoo

Poorhasan Schamchi 1 , Necdet Saglam 2 , Hikmet
Trk Katrcoglu 3 , Nilfer Aksz 4
1 Institute of Science, Department of
Biology/Biotechnology, Hacettepe University,
Ankara, Turkey
2 Institute of Science, Department of Nanotechnology
and Nanomedicine, Hacettepe University, Ankara,
Turkey
3 Faculty of Education, Department of Secondary
Science and Mathematics Education, Biology
Education Programmes, Gazi University, Ankara,
Turkey
4 Faculty of Science, Department of Biology,
Hacettepe University, Ankara, Turkey

E-mail address: kerkan87@gmail.com (K. Erkan).

Quorum sensing (QS), also known as bacterial cell-to-cell communication, is used by pathogenic bacteria in the regulation
of virulence expression, antibiotic resistance factors. According to theory, quorum sensing inhibitors (QSI) generally reduce
pathogenicity of the microorganism. Chlorella vulgaris known as
freshwater micro-algae commonly is used in medicine and industrial applications. In our study, effects of C. vulgaris extracts on
acyl-homoserine lactone-regulated phenotypes of reporter strains
were investigated. C. vulgaris extracts were screened for QS
inhibitors used bioreporter strains; Chromobacterium violaceum
CV026, C. violaceum CV12472 and Pseudomonas aeruginosa PAO1.
Detecting anti-quorum sensing activity were used to disc-diffusion
and agar well diffusion assays. In special media, P. aeruginosa PAO1
motility assay was performed inhibition of swarming. The different extracts of C. vulgaris were observed inhibition of violacein
production in the reporter strains in LB medium. In addition to,
a signicant reduction in swarms was recorded on P. aeruginosa
PAO1.The inhibition of violacein production and swarming motility may be depend on direct or indirect interference to QS. As a
result, C. vulgaris that showed inhibitory activity in the previous
assays also inhibited acyl-homoserine lactone-regulated. This case
may indicate reduction of pathogenicity. C. vulgaris extracts were
screened for its anti-quorum sensing activity by bioreporter strains.
These data suggest that C. vulgaris different extract for the development of anti-quorum sensing molecules, with possible applications
in the control of bacterial diseases.
http://dx.doi.org/10.1016/j.jbiotec.2014.07.336

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Laser interference microscopy of erythrocyte and Raman spectroscopy were used to investigate the inuence of resveratrol
on human blood erythrocytes. Hydrogen peroxide (H2 O2 ) at a
concentration of 105 M was added to initiate the oxidation
processes. Resveratrol preparation (C14 H12 O3 ; M = 128) at concentration 104 M was added to the erythrocytes suspension prior to
H2 O2 introduction. We found that 105 M H2 O2 slightly reduced
such parameters as the area, phase height, phase volume and thickness of erythrocytes. Thus, preincubation with resveratrol caused
signicant changes: decreasing of area on 35.48%, phase height on
39.02%, phase volume on 60.25% and thickness on 39.01%. Moreover, we observed decreased severity of symmetric and asymmetric
vibrations of the pyrrole rings of hemoglobin under the inuence of
H2O2 on 17.77%. In the case of resveratrol this parameters raised to
7.26% under control value. These evidence correlate with the number of extended porphyrin in hemoglobin by the action of H2 O2
(41.62%). Protective effect of resveratrol resulted in reducing the
amount of extended porphyrin on 12.45%. We also observed the
decrease of the level of malondialdehyde under the inuence of
resveratrol (10.64% compared with the H2 O2 samples).
http://dx.doi.org/10.1016/j.jbiotec.2014.07.337
Functional relevance of prion protein in the
regulation of endothelial copper status
Emanuela Urso, Lidia De Riccardis, Antonia
Rizzello, Antonio Danieli, Raffaele Acierno,
Michele Mafa
Department of Environmental and Biological
Sciences and Technologies, Salento University, Lecce,
Italy
E-mail address: michele.mafa@unisalento.it (M. Mafa).
Prion protein (PrPC) is a ubiquitous membrane-anchored glycoprotein, harbouring four high-afnity copper binding motifs, which
strongly imply a protein involvement in the ion cell homeostasis.
The multifaceted PrPCcopper relationship has been widely investigated in a variety of cell types, but little is known about endothelia.
In accordance with broadly shared hypothesis, endothelial prion
protein essentially contributes the spread of vascular networks by
serving as an adhesion protein and participating in the cytoskeletal reorganization of lamellipodia. Herein, the role of PrPC in the
endothelial management of copper distribution has been analysed by silencing the protein expression in HUVECs. Preliminary
investigations evidenced a reduced morphogenetic potential in
PrPC-knockdown cells, in agreement with literature data. Then,
copper uptake assays revealed a reduced ability to absorb ions,
associated with a depressed membrane Cu(II) reductase activity.
In addition, PrPC interactive properties envisioned the involvement of a prototypal heparan sulfate proteoglycan, Glypican-1, in
the co-regulation of copper transport. Stimulation of Glypican-1
endocytosis through the administration of polyamines has been
found to favour copper uptake in PrPC-expressing cells. Conclusively, data suggest that PrPC expression supports the endothelial