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The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
19
REACTIVE OXYGEN SPECIES IN PATHOLOGY.
EXPERIMENTAL AND CLINICAL STUDIES
C l a u d i a B o r z a 1 , R o d i c a M a t e e s c u 2 , E r i k a D e a k 2 , G a b r i e l a N e g r i s a n u 2 , G e r m a i n e S a v o i u 3
1Departament of Physiolgy, 2Department of 3rd Internal Medicine, 3Department of
Pathophysiology, University of Medicine and Pharmacy Victor Babes Timişoara,
Romania, Pta E. Murgu no 2, Mail: Claudia_borza@yahoo.com
ABSTRACT
Objective: It was studied the antioxidant capacity decreasing, expressed by
superoxidedismutase (SOD) and reduced glutathione (GSH) in experimental and clinical
studies. Method: In experimental asthma we used an experimental model with Sprague-
Dawley male rats sensitized with ovalbumin. In clinical study were investigated
asthmatic patients and 40 patients with noninsulin-dependent mellitus diabetes (NIDDM).
Results: SOD - the control value was 10.7 UI/ml and in sensitized group
significantly decreased at 7.6 UI/ml (p<0.01). In asthmatic children group the SOD
value was 9.02 UI/ml, comparatively with 11.1 UI/ml - the control value (p<0.01). GSH -
the control value was 606 uM/l and in sensitized group significantly decreased at 325
uM/l (p<0.001). In asthmatic children group the GSH value was 272.2 uM/l,
comparatively with the control value: 432.2 um/l (p<0.001).In patients with mellitus
diabetes, SOD significantly decreased at 6.8 +/- 1.2 UI/ml (p<0.001, normal value, 10
UI/ml). GSH significantly decreased at 342 +/- 87 μM/l (p<0.001, normal value 600μM/l).
Conclusion: These parameters express the endogenous antioxidant capacity,
significantly decreased in sensitized animals, asthmatic children group and in diabetic
patients in correlation with oxidative stress intensity.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
20
CARBOHIDRATE-DEFICIENT TRANSFERRIN AND
GAMMA-GLUTAMYLTRANSPEPTIDASE IN THE
EVALUATION OF ALCOHOL CONSUMPTION
B . B u m b a c i l a 1 , D a n i e l a I o n e s c u 2 , A d r i a n a K a y c s a 1 , C r i s t i n a D e h e l e a n 2 , E l e n a G a l c a 3
1University of Medicine and Pharmacy ”Victor Babes”, Biochemistry Department
2University of Medicine and Pharmacy ”Victor Babes”, Faculty of Pharmacy, Toxicology Department, E. Murgu no 2, Timisoara, 300041, Romania, daniela_ionescu2002@yahoo.com
3Lugoj Hospital, Department of Psychiatry
ABSTRACT
Aim: The objective of this study was to compare carbohydrate-deficient
transferrin and gamma-glutamyl transpeptidase (GGT) assays for the evaluation of alcohol
consumption. Methods: 165 patients (100 male and 65 female) were included in this
study. Patients were divided into five categories according to alcohol consumption:
category 1 included non-weaned patients drinking more than 30g/day for women and
more than 50 g/day for men, category 2 included relapse patients, category 3 included
moderate drinkers, category 4 included patients weaned less than one month, and
category 5 included patients weaned more than one month. A specifically standardized
questionnaire was used. Results: Sensitivity of CDT varied, depending on patient category,
from 36% to 96% versus 45% to 70% for GGT. Specificity of CDT varied from 72% to 90%
versus 21% to 61% for GGT. After one month of abstinence specificity of CDT was 65%
versus 18% for GGT. Conclusion: This study confirms that carbohydrate-deficient
transferrin is more accurate in predicting alcohol consumption compared with gamma-
glutamyl transpeptidase in alcoholic patients evaluation.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
21
HPLC SEPARATION OF CAROTENOIDS FROM
PHYSALIS ALKEKENGI L. IN DIFFERENT STAGES OF
MATURATION
A n d r e a B u n e a , A d e l a P i n t e a , C a r m e n S o c a c i u
Department of Chemistry and Biochemistry, University of Agricultural Sciences and
Veterinary Medicine, Manastur 3-5, 400372,Cluj-Napoca, Romania;
andrea_bunea@yahoo.com
ABSTRACT
Physalis alkekengi L. is an ornamental plant known as „chinese lantern” which
also produces edible fruits rich in carotenoids. The carotenoid content and profile of
Physalis sepals have been determined in different stages of maturation.
The carotenoid content in all samples was determined comparatively, using
spectrophotometric and HPLC-PDA analysis on RP-18 column. The total carotenoid content
in green sepals was 203.72 mg/kg dry weight, 945.08 mg/kg dry weight in yellow sepals
and 2269.92 mg/kg dry weight in orange ones. The major carotenoid content was found in
orange sepals of Physalis alkekengi L. at full ripeness.
Chromatographic separation of the unsaponified extract by HPLC showed the
presence of β-cryptoxanthin monoesters and zeaxanthin diesters at more than 85% in the
yellow and orange sepals while in green ones they are totally absent. Other carotenoids
found were: neoxanthin, violaxanthin, antheraxanthin and unesterified β-cryptoxanthin
and zeaxanthin. The common carotenoid found in all stages of maturation was β-carotene.
The LC-MS analysis confirmed that β-cryptoxanthin and zeaxanthin are esterified only with
middle chain saturated fatty acids, mainly palmitic acid. The carotenoid esters biosynthesis
starts in yellow sepals and rich the full concentration in the orange sepals. Physalis
alkekengi L. is a valuable source of carotenoids and especially of zeaxanthin.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
22
THE EFFECT OF A PROPOLIS EXTRACT
ADMINISTRATION IN CADMIUM INDUCED
OXIDATIVE STRESS, IN RATS
I u l i a n a C h i s - B u i g a , L u c i a O l a r i u , M i h a e l a P u p
Faculty of Veterinary Medicine, Timişoara 119-Calea Aradului, 300645-Timisoara,
ROMANIA luciaolariu@yahoo.com
ABSTRACT
The experimental oxidative stress due to a single doze administration of cadmium
(20 ppm cadmium as cadmium chloride) and the protective antioxidant action of an
alcoholic propolis extract was studied. The following parameters: malondialdehide,
catalase, superoxid dismutase, glutathione peroxidase, glutathione reductase, glutathione
and the liver and kidney cadmium concentrations were investigated.
The obtained results revealed that a single doze of 20 ppm of cadmium could
generate a significantly increasing of malondialdehide and of the superoxid dismutase
activity. Decreasing of glutathione peroxidase, glutathione reductase and catalase
activities were registered. The cadmium concentration in liver and kidney was significantly
lower after a propolis extract administration.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
23
EVALUATION OF SOD AND GPX ACTIVITIES IN
SCHIZOPHRENIA
E l e n a C r i s t i n a C răc i u n 1 , M a r i a D r o n c a 2 , B o g d a n N e m eş 3 , D o i n a C o z m a n 3 1Pharmaceutical Biochemistry and Clinical Laboratory Department, Faculty of
Pharmacy;
2Medical Biochemistry Department, Faculty of Medicine;
3 Clinical Psichology and Mental Health Department, Faculty of Medicine;
„Iuliu Haţieganu” University of Medicine and Pharmacy Cluj-Napoca
ecgagyi@yahoo.com
ABSTRACT
In recent years there is great evidence that oxygen free radicals play an
important role in the pathophysiology of many neuropsychiatric disorders. In
schizophrenia, antioxidant status could be altered as a consequence of both the evolution
of the disease and the neuroleptic treatment. In the present study we investigated the
activities of erythrocyte superoxide dismutase (SOD) and whole blood glutathione
peroxidase (GPx) in 70 schizophrenic patients and 43 healthy volunteers, age and sex
matched. We have observed significantly higher values of SOD activity (1486.45 ± 262.59
U/g Hb versus 1393.92 ± 250.73 U/gHb, p< 0.05) but normal values of GPx activity
(38.86 ± 9.59 U/g Hb versus 44.24 ± 11.53 U/g Hb, p>0.05) in the schizophrenia group
compared with the control group. These results suggest an adaptative response to the
increased superoxide radicals production.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
24
OXIDATIVE EFFECTS AFTER PHOTODYNAMIC
THERAPY IN RATS
D o i n a D a i c o v i c i u 1 , A d r i a n a F i l i p 1 , S i m o n a C l i c h i c i 1 , S o i m i t a S u c i u 1 , A d r i a n a M u r e s a n 1 , N i c o l e t a D e c e a 1 , S i m i n a D r e v e 2
1Physiology Department, Medicine and Pharmacy University Cluj-Napoca 13 Emil
Isac St., Cluj-Napoca, ROMANIA .(daicoviciu@yahoo.com)
2National R&D Institute of Isotopic and Molecular Technologies, Cluj-Napoca
ABSTRACT
The oxidative effects of photodynamic therapy with 5-aminolevulinic acid (5-ALA)
and chitosan were evaluated in Wistar rats s. c. inoculated with Walker carcinoma. The
animals were irradiated with red light (λ=685 nm, D=50 J/cm2, 15 min) 3h after i.p
administration of 250 mg/kg b.w. 5-ALA or 1.5-mg/kg b.w. chitosan. The lipid peroxides,
protein carbo-nyls, -SH groups and antioxidant capacity (DPPH test) were determined in
blood and tu-mor tissues. 24h after the treatment, in 5-ALA treated animals the level of
lipid peroxides in tumors was increased as compared to controls (0.75±0.24 vs. 0.38±0.09
nmoles MDA/mg protein, p<0.05). Protein carbonyls had a higher level in tumor tissues of
5-ALA group comparatively to controls and chitosan group (6.18±3.00 vs 1.55±0.42 and
1.92±0.09 nmoles/mg protein). Chitosan had an antioxidant effect expressed by lipid per-
oxides level (0.22±0.09 nmoles MDA/mg protein). Our results suggest that photodynamic
therapy is associated with biomolecules alterations produced by oxygen reactive species.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
25
IN VITRO AND IN VIVO TESTS TO EVALUATE OF THE
EFFICACY AND TOXICITY OF BETULINIC ACID
STANDALONE OR MIXED WITH HPGCD
C r i s t i n a A . D e h e l e a n 1 , C . Ta t u 1 , C o d r u ţa Şo i c a 1 , A l e x a n d r a G r u i a 1 , V. O r d o d i 1 , S . C î n tă - P î n z a r u 2 , A d r i a n a K a y c s a 1
1University of Medicine and Pharmacy „Victor Babeş” Timişoara, Square Eftimie Murgu no.2, 300041, Timişoara, e-mail cadehelean@umft.ro 2Babes Bolyai University, Faculty of Physics, Molecular Spectroscopy Department, Kogalniceanu 1, RO 400084 Cluj-Napoca, Romania
REZUMAT
Betulinic acid is a naturally occurring pentacyclic triterpene, intensive analysed for
its anticancer selective effects on some tumour cells (e.g., melanoma cells). Its
mechanism of action consists in inducing selective apoptosis. The objective of the present
study was to analyze the antitumour effect of betulinic acid standalone or mixed with
HPGCD (hydroxypropyl gamma cyclodextrin) as antileukaemia agent. Its activity was
determined on K562 erythroleukaemia cell line. The antitumour activity was also analyzed
from the angiogenic activity point of view. The toxic activity was appreciated on human
mesenchymal stem cells. In vivo tests consisted in the analysis of the betulinic acid
solution, starting from 12,5 mg/ml on embryonated egg model. Materials and methods
were specific for the in vitro and in vivo evaluations: MTT assay, mycroscopic evaluation
plus spectral analysis. The dissolution of the compounds in an aqueous environment
(serum) was achieved by adding of HPGCD 1:2 ratio as was indicated in literature data.
DMSO was used for dissolution for in vitro tests. The conclusions are that betulinic acid is
active in low doses on the tested tumour cells and has a reduce toxicity. It induces a dose-
dependent anti-angiogenic effect on the embryonated egg.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
26
ANTIOXIDANT PROTECTION IN AUTISM
M . D r o n c a 1 , E . C . C răc i u n 2 , S . P. P a s c a 1 , F. I f t e n e 1 , I . B e n g a 1 1Faculty of Medicine; 2Faculty of Pharmacy, „Iuliu Haţieganu” University of Medicine
and Pharmacy Cluj-Napoca (m_dronca@yahoo.com )
ABSTRACT
Autism is a severe neurodevelopmental disorder defined by social and
communication impairments and stereotyped behaviors. The underlying etiology of autism
is unknown. It is suggested that autism may result from an interaction between genetic,
environmental, and immunological factors, with oxidative stress as a mechanism linking
these risk factors. Knowing that the brain is highly vulnerable to oxidative stress due to its
limited antioxidant capacity and higher amounts of lipids and iron, we investigated the
activity of the erythrocyte superoxide dismutase (SOD) and whole blood glutathione
peroxidase (GPx) in patients with typical and atypical autism. We investigated 41 children,
21 with typical autism (age 4.82 ± 0.33 years) and 20 with atypical autism (age 8,99 ±
0.81 years) followed at the Pediatric Psychiatry and Pediatric Neurology Clinics in Cluj-
Napoca. The control children were free of any neuropsychiatric disorder, age and sex
matched. We observed normal level of GPx activity in both typical (44.66 ± 2.92 U/g Hb)
and atypical (42.02 ± 2.34 U/g Hb) autistics in comparison with age matched controls
(47.63 ± 3.30 U/g Hb and 44.23 ± 2.74 U/g Hb respectively). On the other hand, the
activity of SOD was significantly decreased only in children with atypical autism compared
with matched age controls (1154.28 ± 79.62 U/g Hb versus 1395.10 ± 60.18 U/g Hb, p<
0.05). In conclusion, our results indicate that the antioxidant defense is altered in atypical
autism, but not in typical autism.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
27
THE TOTAL ANTIOXIDANT STATUS IN
HYPOTHYROIDISM
D . F i r u , S t e f a n a P o p e s c u , D a n i e l a Ta c h e , A n a K i e s e
Str. Petru Rares, no. 4, 200349, Craiova, Dolj stoapo@yahoo.com
ABSTRACT
The importance of the iodine ion oxidation to elementary iodine during
thyroidian hormone biosynthesis was well established . The iodine ion deficiency may
produce oxidative stress conditions by the increase of H2O2 concentration .The iodine
excess of thyroidian cells determines cytotoxicity dependent on the formation of free
radicals .These cytotoxic effects may depend on the total antioxidant status. In our
reaserch we watched the oxidative stress by lipidic peroxide products – malonil dialdehida
(MDA)-and the total antioxidant status for a group of hypothyroidian patients in the
Endocrinology Clinic of The University Hospital of Craiova .
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
28
THE INFLUENCE OF CULTIVATION CONDITIONS
UPON COQ PRODUCTION BY YEASTS
L u m i n i t a G e o r g e s c u a n d A n c a N i c o l a u
University „Dunarea de Jos” Galati, 47 Domneasca St., 800008 Galati,
luminita.georgescu@ugal.ro and anca.nicolau@ugal.ro
ABSTRACT
Among microorganisms, yeasts are renamed for producing compounds from the
coenzyme Q group, but their use as CoQ10 industrial producers is not very popular
because of the bioprocess cost. In spite of this, natural CoQ is prefered to the synthetic
one as supplement in medicine, food and cosmetic products. In an attempt to improve the
CoQ10 bioproduction, three yeasts were investigated in our laboratory. The yeasts used in
the experiments were Saccharomyces cerevisiae from Pakmaya, a Candida sp. isolated
from wine and a Rhodotorula sp. isolated from fruits. The yeasts were cultivated on a
minimal medium based on sucrose and amonium salts, both under stationary and shaked
conditions in order to evaluate the importance of aeration in CoQ biosynthesis. Then, the
influence of different substances (hydrogen peroxide, cysteine, carrot juice, molasses)
upon CoQ10 production was tested. In order to extract the CoQ, yeast biomass was
submitted to repeated cycles of freezing-thawing. Coenzyme Q quantified by HPLC in n-
propanol extracts was better synthetised by Rhodotorula sp. Reverse-phase HPLC was
performed on an Agilent 1200 series system with a multiple wavelength detector under
isocratic conditions. The separation of CoQ was performed on a PRP-1 column with a
mixture of acetonitrile and tetrahydrofuran as mobile phase. CoQ retention time was 3.97
at 1 ml/min flow rate and 30°C.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
29
THE DEPENDENCE OF ANTIOXIDANT ACTIVITY SEA
BUCKTHORN OILS UPON FATTY ACIDS
COMPOSITION
A l e x a n d r a T. G r u i a 1 , 2 , M a r i a n a A l b u l e s c u 1 , C a l i n A . Ta t u 2 , V a s i l e O s t a f e 1 1West University of Timisoara, Faculty of Chemistry-Biology-Geography, Department
of Chemistry, 16 Pestalozzi, Timisoara, 300115, Romania
2Transplant Immunology Department, Clinical Laboratory, County Hospital, Bd. Iosif
Bulbuca No.156, 300736 Timisoara, Romania
ABSTRACT
Sea buckthorn plants contain an array of nutrients with antioxidant activity, used
in medicinal purposes, as carotenoids, vitamins, phytosterols, flavonoids, etc. By its rich
content in essential fatty acids, different parts of berries are used to obtain the oil. Due to
its powerful antioxidant effects, the oil has topical applications in medicine. The aim of the
study was to establish the content of fatty acids from two different sea buckthorn oils
pharmaceutical products, and to evaluate their antioxidant capacity. In order to perform
the analysis, the samples were derivatized prior to injection in gas-chromatograph (GC)
and the compounds were identified with mass spectrometry (MS). The antioxidant activity
was evaluated with 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Analysis of phases,
estherified and saponified, of sea buckthorn oils revealed a difference in the content and
structure of fatty acids from assayed samples. Comparing the results, one can establish
which oil is richer in polyunsaturated fatty acids and thus, better in radical scavenging
activity.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
30
IRON SUPPLEMENTATION IN HEMODIALYSIS -
INFLUENCE ON OXIDATIVE STRESS PARAMETERS
M a n o l e s c u B , S t o i a n I , C a p u s a C , M i r c e s c u G , L i x a n d r u D , A t a n a s i u V, C o l t a n L , G a m a n L
University of Medicine and Pharmacy "Carol Davila", Bucharest
ABSTRACT
Oxidative stress in hemodialysis is considered to be the result of metabolic
changes in chronic renal failure, loss of low weight antioxidants during hemodialysis
session, biological incompatibility of the dialitic system and medication (especially
parenteral iron used to treat renal anemia). Iron administration can amplify oxidative
stress, most of the pharmaceutical preparations having loosely bound iron. From a
theoretically point of view arterial iron administration can be less harmful compared with
currently clinical venous administration. We have considered of interest to study the
influence of the two different routes of iron administration on erythrocyte antioxidants.
Superoxide dismutase, catalase, glutathione peroxidase and transferase activity and the
level of total and non proteic thiols were analyzed on 20 hemodialised patients receiving
parenteral iron therapy. In order to gain information on others antioxidants present in
these cells we have also determined TEAC on erythrocytes lysates. There were no
statistically significant differences between the levels of these parameters when iron was
administered through the arterial or venous route with the exception of non-proteic thiols
and TEAC wich were higher for arterial iron administration suggesting a possible benefit of
this route In order to definitely clarify if there is or there is not a marked difference for the
arterial versus venous iron administration in these patients we need to study also the
plasmatic variations of oxidative stress parameters.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
31
MODIFICATION OF HSP 70 AND HSP 27 EXPRESSION
AND OXIDATIVE STRESS IN MANGANESE TREATED
HUH7 CELLS
M u n t e a n u M a r i a C r i s t i n a , P o s t o l a c h e L a u r a , B o d e a L . , C o s t a c h e M a r i e t a , D i n i s c h i o t u A n c a
University of Bucharest, Biochemistry and Molecular Biology Research Center, Spl. Independentei, 91-95, Bucharest, Romania, dinischiotu@yahoo.com; cristina.munteanu@bio.unibuc.ro;
ABSTRACT
High levels of manganese are toxic for a number of tissues such as liver, brain, gills and different cell cultures. Prolonged Mn(II) exposure induces the formation of reactive oxygen species leading to impairment of the antioxidant system.The aim of the present study was to evaluate the HUH7 cells adaptative response to experimental manganese exposure by biochemical and molecular methods, with particular attention to oxidative metabolism and heat shock protein gene expression. Antioxidative enzymes (catalase, superoxide dismutase) changed their profiles after 12 and 18 hours of 5mMMn(II) treatment. The total SOD specific activity was increased possibly because of a higher level of generated superoxide anion and of increase in Mn–SOD expression that was accompanied by an elevation of its activity. Hsp27 and Hsp 70 protect the cells against toxic effects triggered by a variety of stimuli including oxidative stress. More specifically, it was shown that Hsp 27 reduced the production of reactive oxygen species. It seems that this protein could modulate the intracellular redox state. Using Real Time PCR, we have shown a significantly increased Hsp 27 mRNA expression after 2 hours of treatment with 5mM Mn(II) on Huh7 cells, whereas the level of Hsp70-1B mRNA increased immediately after the exposure (0.5 hr). Our results revealed that, the transcription of hsp70-1B and hsp27 at high levels occurred in the first 2 h after manganese chloride treatment. Later the transcription rate decrease, but is higher for hsp 70-1B than hsp27.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
32
P53 AND RB (RETINOBLASTOMA) GENE
EXPRESSION IN TUMORAL CELLS, UNDER THE
INFLUENCE OF VITAMIN E
A . - M . N i c u l e s c u ( 1 ) , L . G . G h e t e a ( 1 ) , R . M o t o c ( 1 ) , G r. M i h a e s c u ( 1 ) , R . H u c u l e c i ( 2 ) , C . D i a c o n u ( 3 ) , C . U r s a c i u c ( 4 ) , G . S a v i ( 4 )
(1)Institute of Genetics, University of Bucharest, Romania, (2) Multiple Users Research Base – Molecular Biology, University of Bucharest, Romania, (3) Institute of Virology “Stefan S. Nicolau”, Bucharest, Romania, (4) National Institute for Research & Development in Pathology and Biomedical Sciences “V.Babes”, Bucharest, Romania, niculescuana2006@yahoo.com
ABSTRACT
The aim of this study was to establish the in vitro and in vivo effects of vitamin E
(vit E) on Wistar rat Walker sarcoma 256 ascitic cells. The treatments with vit E were
administrated in different doses and time intervals. The analyses were performed by flow
cytometry, for the study of apoptosis dynamics, and by Real-Time PCR, for p53 and rb
gene level expression detection. The results obtained revealed that in vitro treatment with
vit E had the most significant result on the number of apoptotic cells after 48 hours of
treatment, when the maximum percentage of apoptosis was obtained (24,14%), while the
control culture presented a percentage of 3.45% of apoptotic cells. Real-Time PCR analysis
showed that the expression level increased for both genes, at two concentrations of the vit
E used in the experiments, indicating a increased transcription activity for p53 and rb. The
most significant result, for the in vivo experiments, was an increase of the life span of the
rats treated with vit E, but, in this case, none of the two genes (p53 and rb) level was
modified. This results suggest that, in vivo, vit E could have a stimulating effect mainly on
the immune system which become more efficient in decreasing the tumor progression
rate.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
33
STUDY OF DEUTERIUM DEPLETED WATER EFFECT
IN EXPERIMENTAL CADMIUM CHLORIDE INDUCED
OXIDATIVE STRESS IN FEMALE RATS
L u c i a O l a r i u , M i h a e l a P e t c u , I u l i a n a C h iş - B u i g a , C a m e l i a T u l c a n , M i h a e l a P u p , I l e a n a B r u d i u
Faculty Of Veterinary Medicine, Timişoara, 119-Calea Aradului, 300645-Timisoara,
ROMANIA (luciaolariu@yahoo.com)
ABSTRACT
The present work deals with the study of the deuterium depleted water (DDW)
effect on female rats with cadmium induced oxidative damages (20 ppm Cd/ b.w. in single
dose administration). The deuterium concentration in surface water is about 150 ppm
(over 16 mM) and more than 10 mM in living organisms. The possible role of the naturally
occurring deuterium in biological systems was not studied before 1993, when the first
experiments with DDW revealed that due to deuterium depletion the non-tumorous L292
fibroblast cells required longer time to multiply in vitro and DDW caused human breast
tumor regression in mice. In our study there was determined that in a short time
treatment (30 days) DDW has a prooxidant effect (malondialdehide average values are
increasing, glutathione are decreasing); the antioxidant system response was evident.
After Cd intoxication and 61 days treatment the superoxid dismutase and glutathione
reductase activities were stimulated. The glutathione-peroxidase registered decreased
activity values as controls.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
34
IN VITRO ANTICANCER ACTIVITY OF THE EXTRACT
OF HORSE CHESTNUT FOLIAR BUDS, AESCULUS
HIPPOCASTANUM L.
C a m e l i a P e e v 1 , C r i s t i n a D e h e l e a n 1 , A d r i a n a K a y c s a 2 , C o d r u t a S o i c a 1 , 1University of Medicine and Pharmacy “Victor Babes”, Faculty of Pharmacy, P-ta
Eftimie Murgu No.2, Timisoara 300041, Romania, cameliapeev@yahoo.com
2 University of Medicine and Pharmacy “Victor Babes”, Department of Biochemistry,
P-ta Eftimie Murgu No.2, Timisoara 300041, Romania
REZUMAT
The horse chestnut foliar buds are a new category of vegetal products that are
used in in the modern phytotherapy, also called gemmotherapy. The natural compounds,
present in small quantities, develop a synergic anticancer activity. In this study we have
determined the water content, total nitrogen, proteins, hemolytic saponins and the
presence of esculin by TLC. The anticancer activity of the crude extract was demonstrated
by the comparative biotest on Lepidium sativum, and, in vitro, on neuroectodermal cell
types from skin carcinoma (A431) and breast cancer cells (MCF 7).
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
35
FUNCTIONAL ANALYSIS OF FOUR SELECTED
C/EBPα GENE PROMOTER POLYMORPHISMS
PREVIOUSLY ASSOCIATED WITH FEATURES OF THE
INSULIN RESISTANCE SYNDROME/T2D
C o r i n a S a m o i lă , L i v i u Tăm aş , E d w a r d Şe c l a m a n , A d r i a n a N e g h i nă Biochemistry Department, University of Medicine and Pharmacy “Victor Babes”, Timisoara, biochim@umft.ro
ABSTRACT
The CCAAT / enhancer binding protein alpha, CEBPα, is a member of CEBP family
transcription factors. All members (C/EBPα-C/EBPζ) contain basic leucine zipper domain
(b-ZIP), in the C-terminus region, involved in DNA binding and dimerisation. Multiple roles
of the C/EBP family were identified in the control of cell cycle and differentiation processes,
in metabolism and inflammation. Studies have demonstrated that the CEBPα plays a key
role in the process of adipocytes differentiation. A new interest has been developed in the
analysis of associations of the CEBPα with the development of the insulin resistance
syndrome/T2D. Several common polymorphic variants of CEBPα promoter associated with
haemeostatic and metabolic factors related to the insulin resistance syndrome/T2D were
previously identified in a region spanning 3 kb from CEBPα promoter. In this study, using
stable transfections of the 3T3-442A cells (mouse preadipocyte cell line) with polymorphic
and wild type promoter constructs, we measured activities of CEBPα promoters to asses
the effects of four haplotype constructs on gene transcription in order to identify functional
sites important for regulating the level of CEBPα gene transcription in 3T3-F442A CEBPα-
expressing cell line. Analyses of a series of promoter constructs containing the four gene
haplotypes, either in preadipocytes or differentiated adipocytes, showed significant higher
activity of one promoter construct comparative with the others.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
36
TRANGENOMIC WAVE™ DNA FRAGMENT ANALYSIS
OF A C/EBPβ GENE PROMOTER FRAGMENT
C o r i n a S a m o i lă a , A n d r e i A n g h e l a , A l f a X e n i a L u p e a b , M a r i l e n a M o t o c a a Biochemistry Department, University of Medicine and Pharmacy “Victor Babes”,
Timisoara, biochim@umft.ro
bBiochemistry Department, University of Polytechnics, Faculty of Engineering and
Environmental, Timisoara
ABSTRACT
Functional analysis of human promoter provides information about the gene
regulation mechanisms and the molecular networks involved. In this study, we have
investigated the association of C/EBPβ and atherosclerosis, using Trangenomic WAVE™
DNA Fragment Analysis on a fragment of 248bp from C/EBPβ gene promoter, to identify
the gene promoter polymorphisms possible associated with atherosclerosis and other
cardiovascular risks factors in 144 subjects with significant atheroma (1,2 or 3 major
coronary arteries with >50% obstruction, and/or peripheral artery lesions and/or at least
one atheromatous carotid stenosis demonstrated by angiography and echography) and in
150 subjects with no significant atheroma. Analysis revealed two adjacent polymorphisms
-411/-412 CG /TT. No heterozygous individuals for each single polymorphism were
distinguished. There was no difference in the genotype distributions of patients with
significant atheroma (haplotypes frequencies: -411/-412 CG [30.5%], -411/-412 CGTT
[50.7%], -411/-412 TT [18.8%]) and without significant atheroma (haplotypes
frequencies: -411/-412 CG [28.9 %], -411/-412 CGTT [48.3%],-411/-412 TT[22.8%]).
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
37
DIABETES VS CHRONIC RENAL FAILURE -
OXIDATIVE AND CARBONYL STRESS
S t o i a n I . , C a p u s a C . , M i r c e s c u G . , A t a n a s i u , V. , M a r a c i n e , M . , V i r g o l i c i , B . , M u s c u r e l C . , L i x a n d r u D . , M a n o l e s c u B . , G a m a n L .
University of Medicine and Pharmacy "Carol Davila", Bucharest
ABSTRACT
Advanced glycation end products (AGE) have been found increased in blood
samples from both diabetes and chronic renal failure patients suggesting a relationship
between carbonyl stress (accumulation of substrates due to excess production or impaired
clearance) and oxidative stress. Because both conditions have similarities regarding the
presence of AGE but different causes for this presence we have considered of interest to
study parameters of carbonyl and oxidative stress in diabetic and chronic renal failure
patients.We have selected 18 well controlled type II diabetes patients, 20 hemodialysed
patients with diabetes, 20 hemodialysed patients not having diabetes and 22 age matched
control subjects. On fasting blood samples we have determined routine biochemical
parameters as well as dicarbonyl compounds, Amadori Products (precursor of AGE),
TBAR'S, TEAC, antioxidant "gap" and total plasmatic thiols. In diabetes patients the
presence of oxidative stress is suggested by increased TBAR'S. In these patients there is
no difference from the controls for the level of TEAC antioxidant "gap" and total thilos. The
carbonyl stress markers studied (dicarbonyl compounds and Amadori Products) are also
not statistically significant different from the controls. For chronic renal failure patients
with end renal disease treated by hemodyalisis, with or without diabetes, we have put in
evidence the presence of both oxidative and carbonyl stress, the parameters studied being
significantly different from the controls, the patients suffering from both diabetes and
chronic renal failure looking the most vulnerable.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
38
THE BIOMEDICAL APPLICATIONS OF THE MAGNETIC
NANOPARTICLES
M a r i a n a Ş i n c a i , D i a n a A r g h e r i e , D i a n a G â n gă , D o i n a B i c a
Faculty of Veterinary Medicine, Timişoara, 119-Calea Aradului, 300645-Timisoara,
ROMANIA, (amsincai@yahoo.com)
ABSTRACT
Our researches about the biomedical applications of the magnetic nanoparticles
were directed in three domains. For the cicatrizing and anti-inflammatory effects
were experimented three types of magnetic fluids on skin lesions at guinea pigs, mice and
dogs and some volunteer persons. Comparatively, we used an existing pharmaceutical
product on control skin lesions. Periodically, from the experimental group animals, we
gather small skin fragments for histological study. Both the clinical and the histological
exams pointed out that the magnetic nanoparticles had an anti-inflammatory and rapid
cicatrizing effect. For the antitumoral effect of the biocompatible magnetic fluids, the
experiments were experimented on bitch and cats mammary tumors and fibro sarcoma.
The best results were obtained using the magnetic fluid based on water with magnetite
nanoparticles and with the magnetic fluid based on water with cobalt ferrite nanoparticles.
In all studied cases, we observed that in the first week the evolution of the tumors was
stopped. After that, step by step, the tumors reduced in size. To point out the UV-
protector effect were experimented three types of magnetic nanocompounds on white
mice auricles. The experimental animals were exposed at an UV lamp. After 8 hours it was
observed that the mice auricles protected with nanocompounds was well protected by UV-
radiation comparatively with control auricles at which appeared a severe erythema.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
39
SOD ACTIVITY IN FRESH AND THAWED SEMINAL
PLASMA AFTER CRYOPRESERVATION USING TWO
DILUENTS
T u l c a n C . 1 , C s e h S . 2 , C h i s u I . 1 , O l a r i u L . 1 , I g n a V. 1 , C e r n e s c u H . 1 1Faculty of Veterinary Medicine, 119 Calea Aradului, 300645 Timisoara , e-mail:
camelia_tulcan@yahoo.com
2SzIU, Faculty of Veterinary Science Budapest, Hungary
ABSTRACT
The objective of this study was to characterize superoxide dismutase activity
associated to cryopreservation and its implications on the quality of the dog semen
cryopreserved with two different diluents: Biladyl and Bioxcell.
Twenty-five ejaculates collected from 8 males aged from 1 to 2½ years, were
used in this study. The cryopreservation was performed using the Biladyl and Bioxcell
protocols. The SOD activity was evaluated in fresh and cryopreserved seminal plasma by
NBT method. The quality parameters of semen (motility, viability and morphology) were
evaluated using WHO techniques.
In cryopreservation process, a decrease of the SOD activity is recorded,
compared to that of the fresh seminal plasma. The release of the intracellular SOD and the
onset of the oxidative stress are evidenced in both cases. The fact that the injury level of
the spermatozoa plasmatic membrane integrity has a decisive role on the level of the SOD
activity after thawing is demonstrated in the case of Bioxcell diluter, where the worse
quality of semen was registered.
The Annual International Conference of the Romanian Society of Biochemistry and Molecular Biology
TIMIŞOARA, September 6-8 , 2007
Posters
40
SPECTROPHOTOMETRIC STUDY ON THE
RIBOFLAVIN − PLANT EXTRACTS INTERACTION
M a r i a n a V o i c e s c u 1 , L i l i a n a L u n g u 2 , V a s i l e D i n o i u 2 , M a r i l e n a V a s i l e s c u 1
1Romanian Academy, Institute of Physical Chemistry „Ilie Murgulescu”, Splaiul Independentei 202, 060021 Bucharest, Romania (voicescu@icf.ro) 2Romanian Academy Institute of Organic Chemistry „C.D.Nenitescu”, Splaiul Independentei 202B, 060021 Bucharest, Romania (llungu_cco@yahoo.com)
ABSTRACT
Intrinsic radical scavenging processes of compounds administered with food
have to warrant metabolic control within certain limits. Antioxidants which in many cases
are free radical scavengers or quenchers of activated states comprise a wealth of classes
of organic molecules including phenolics, probably as the most prominent ones. Using
UV-VIS Absoption and Fluorescence Spectroscopy, in this paper mechanism of protection
from oxidative damage of some plant extracts are discussed. Moreover, examples of
antioxidative functions of a few important plant extracts in certain diseases are reported.
Riboflavin - Plant Extracts Interaction is investigated in order to get information if
riboflavin acts as additional sensitizer to the superoxide generation. The results are
discussed with relevance to the oxidative stress process that takes place in humans.
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