Andronikashvili Institute of Physics Department of Physics of Biological Systems nelly_sapojnikova@hotmail.com H1. - презентация

1 Andronikashvili Institute of Physics Department of Physics of Biological Systems H1

2 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

3 HMGB1 Box AHMGB1 Box B HMGB1 Box A (positively charged) C- terminus (negatively charged) HMGB1 Box B (positively charged) The double life of HMGB1 chromatin protein: Architectural factor and extracellular signal HMGB1 is ubiquitous and only 10 times less abundant than core histones. The cytokine-inducing activity resides in the B Box domain The A Box antagonizes HMGB1- induced inflammation

4 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???)

5 Growth Inhibition (% of control) 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). 2 h Cr(VI)+46 h 24 h Cr(VI) 48 h Cr(VI) 24 h Cr(VI) +24 h Concentration of Cr(VI) (μM) Dr Marina Abuladze

6 % of Control ROS (DCFH-DA) ROS (DHE) 2 h 24 h 2 h 24 h AB ROS level in control untreated and Cr(VI)-treated HLF cells

7 GPx activity (mU/ml) GR activity (mU/ml) 2 h 4 h 24 h 48 h A B Dr Nino Asatiani

8 SOD catalyzes O 2 dismutation to H 2 O 2 15 μM Cr(VI)30 μM Cr(VI) C 2h 5 μM Cr(VI) Mn-SOD Cu,Zn-SOD C 2h C 24 h Peak Density (% of Control) Mn-SOD Cu,Zn-SOD 2 h 24 h 2 h 24 h Mn-SOD Cu,Zn-SOD 2 h 24 h Mn-SOD Cu,Zn-SOD A B

9 2 h 4 h 24 h 48 h Catalase activity (U/ml) Dr Tamar Kartvelishvili

10 The time course of the ESR signal intensity in the model Fenton reaction. The model Fenton reaction mixture contained 100 mM DMPO, 1 mM FeSO 4, 100 mM H 2 O 2, 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 mg/ml.

11 Study of the antioxidant system status and blood metalloproteinases cross influence at acute ischemic stroke 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

12 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

13 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

14 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