24 h Cr(VI)

Andronikashvili Institute of Physics
Department of Physics of Biological Systems
[email protected]
H1
Ac-Leu-Ala-His-Tyr-Asn-Lys-Amid (80-85 fragment of H2B) Cu(II) and Ni(II)
Ac-Thr-Tyr-Thr-Glu-His-Ala-Amid (71-76 fragment of H4) Ni(II)
Ac-Glu-Ser-His-His-Amid (C-terminus fragment of H2A) Cu(II) and Ni(II)
Zinc is a novel intracellular second messenger like Ca 2+ and cAMP
HMGB1 is
ubiquitous and only
10 times less
abundant than core
histones.
HMGB1 Box A
HMGB1 Box B
(positively
charged)
(positively
charged)
HMGB1 Box A
The A Box antagonizes
HMGB1- induced
inflammation
The double life of HMGB1
chromatin protein:
Architectural factor and
extracellular signal
C- terminus (negatively charged)
HMGB1 Box B
The cytokine-inducing
activity resides in the B
Box domain
Schematic illustration of potential pathways for HMGB1 release leading to inflammatory responses. HMGB1 can be
exracellularly released by passive secretion from any necrotic cell or by active secretion from activated
macrophages/monocytes.
HMGB1 and diseases: Sepsis; Lung inflammation; Arthritis; Ischemic stroke; Tuberculosis -Mycobacterial infection
induces the secretion of high-mobility group box 1 protein
Bactericidal activity of HMGB1 - functionally belongs to the growing family of antibiotic peptides (Box A???)
Growth Inhibition (% of control)
 2 h Cr(VI)+46 h
Dr Marina Abuladze
 24 h Cr(VI)
 48 h Cr(VI)
 24 h Cr(VI) +24 h
Concentration of Cr(VI) (μM)
Time- and dose-dependent Cr(VI)-induced cytotoxicity by Cr(VI). Fetal human lung
fibroblasts were grown up to 80% confluency prior to chromium treatment at the
concentration range from 2-30 (μM).
A
B
ROS (DHE)
ROS (DCFH-DA)
5 uM Cr(VI)
15 uM Cr(VI)
30 uM Cr(VI)
Control
400
200
350
180
% of Control
% of Control
Control
300
250
200
150
100
5 uM Cr(VI)
15 uM Cr(VI)
30 uM Cr(VI)
160
140
120
100
80
60
40
50
20
0
0
2h
24 h
2h
ROS level in control untreated and Cr(VI)-treated HLF cells
24 h
GPx activity (mU/ml)
160
5 uM Cr(VI)
15 uM Cr(VI)
30 uM Cr(VI)
A
140
120
100
80
60
40
20
0
Dr Nino Asatiani
2h
GR activity (mU/ml)
Control
4h
24 h
48 h
Control
5 uM Cr(VI)
15 uM Cr(VI)
30 uM Cr(VI)
80
70
B
60
50
40
30
20
10
0
2h
4h
24 h
48 h
SOD catalyzes O2
dismutation to H2O2
A
5 μM Cr(VI)
15 μM Cr(VI)
30 μM Cr(VI)
Mn-SOD
Cu,Zn-SOD
C
2h
C
24 h
C
2h
C
24 h
C
2h
B
Peak Density (% of Control)
Control
Mn-SOD Cu,Zn-SOD
180
160
160
140
140
24 h
Control
Cr(VI)
200
200
200
180
Control
Cr(VI)
Mn-SOD Cu,Zn-SOD
180
100
100
100
80
80
80
60
60
60
40
40
40
20
20
20
0
0
0
Mn-SOD Cu,Zn-SOD
Cr(VI)
Mn-SOD Cu,Zn-SOD
2h
Mn-SOD Cu,Zn-SOD
24 h
140
120
2h
24 h
160
24 h
120
120
C
2h
Mn-SOD Cu,Zn-SOD
Catalase activity (U/ml)
Control
5 uM Cr(VI)
15 uM Cr(VI)
30 uM Cr(VI)
160
140
120
100
Dr Tamar Kartvelishvili
80
60
40
20
0
2h
4h
24 h
48 h
The time course of the
ESR signal intensity in the
model Fenton reaction.
The model Fenton
reaction mixture contained
100 mM DMPO, 1 mM
FeSO4, 100 mM H2O2, 50
mM sodium/potassium
buffer pH 7.4 in a final
volume 62.5 l.
The time course of the ESR signal intensity in the model Fenton reaction
in the presence of crude cell extracts of Cr(VI)-treated and untreated
control L-41 cells. (A) - 2 M Cr(VI) treatment; (B) - 20 M Cr(VI)
treatment. - Control;  - 24 h of the cell growth under Cr(VI) action; 
- 48 h of the cell growth under Cr(VI) action. Cr(VI) as potassium
chromate was added to the cell culture at the 48 h of growth (80% of
confluence). Protein concentration in ESR sample was 0.168 mg/ml.
Study of the antioxidant system status and blood metalloproteinases
cross influence at acute ischemic stroke
Zn
Recently much research interest is focused on predictors
of functional outcome in stroke. The development of
early prognostic biochemical parameters is of great
importance. Accumulating data suggest that MMPs have
a deleterious role in stroke. The hypothesis to be tested in
this project is that the profile of plasma matrix
metalloproteinases MMP-2, MMP-9 and MMP-12 is
changed (elevated) during the hyperacute stage of the
disease, and it provides prognostic information for stroke
severity, especially malignant middle cerebral artery
infarct along with other inflammatory markers (plasma
status of antioxidant defense system).
1.To assess quantitatively the level of MMP-2, MMP-9
and MMP-12 in the blood plasma of stroke patients in
the first 24 hours after stroke.
2. To estimate the oxidant/antioxidant balance in the
blood plasma of patients and healthy volunteers and
develop it as the marker of the disease.
3. To measure the blood level of extracellular cytokine
HMGB1 as the possible factor upregulating MMPs in
human stroke.
4. To estimate the predictive role of the above-mentioned
serological markers in development of malignant cerebral
infarcts.
5. To estimate the predictive role of the above-mentioned
serological markers in functional outcome after IS.
3D Structure of MMP
structures - ProMMP-2-TIMP-2
complex.
Orange - propeptide
Green - catalitic domail
Pink -fibronectin domain
Blue - TIMP-2
One catalitic Zn;
one structural Zn;
Three calcium ions
Zn
Биочипы как диагностическое средство для быстрого
скрининга болезнетворных бактерий и вирусов
(кишечный тракт, уро-репродуктивный тракт)
Микрочип - это пластинка, покрытая
тонким слоем фиксирующего
нейтрального вещества (например
модифицированного акриламида), в
которое в совершенно определенной
последовательности нанесены
фрагменты ДНК (олигонуклеотиды),
специфически сконструированные и
синтезированные для диагностики.
•Нуклеиновые кислоты (из
ассоциированных со слизистой облочкой
СРБ из биопсийного материала в случае
больных ЯК и из крови в случае анализа
TORCH-инфекции) выделяются и метятся
флуоресцентными метками
Какой новый продукт / услуга предлагается?
1) микрочип-ЯК для быстрой и достоверной
диагностики бактериального происхождения
язвенных колитов (ЯК) и для наблюдения за
проходящим курсом лечения антибиотиками;
Больные и здоровые люди являются
носителями сульфат редуцирующих бактерий
(СРБ), однако профиль бактериальных
популяций обнаруживает поразительное
различие.
АНАЛОГА НЕТ
Какой новый продукт / услуга предлагается?
2) микрочип-TORCH, который позволят с
малыми затратами, быстро провести
мониторинг TORCH-инфекции на первых
месяцах беременности. Этот мониторинг может
стать повсеместным необходимым и
обязательным анализом.
TORCH-инфекция - это комплекс инфекций:
Toxoplasma Gondii, Rubella, Cytomegalovirus,
Chlamidia trachomatis, Herpes virus
Для мониторинга TORCH-инфекции, существует два диагностических
метода