1. Main Topics of Research in the Health Part Effects of garlic powders and known compounds on cellular functions related to arteriosclerosis or inflammation Univ. Munich / Univ. Leipzig / TNO The Netherlands Bioavailability and metabolic fate of garlic compounds INRA Dijon / Lichtwer Berlin Search for new effectors in garlic powder Univ. Leipzig

2. Activation of the transcription factor NF-kB Garlic constituents showed no inhibition of TNF  induced NF-  B activation. Only Printanor 0 solved in Media was able to inhibit TNF  induced NF-  B activation. (Univ. Munich / measured by a new expression assay) Results:

3. Effects of garlic extracts or compounds on liberation of cytokines from human blood cells Garlic extracts are able to reduce LPS induced liberation of the pro-inflammatory cytokines Il-1  and TNF . Garlic extracts are able to increase the LPS induced liberation of anti-inflammatory cytokine Il-10. It is noticeable that unfertilised garlic is less active than fertilised garlic. The liberation of cytokines through RPMI garlic extracts is most likely mediated by endotoxin contaminations.

4. Il-1 

5. Inhibition of MMP-Production by garlic compounds (Univ. Leipzig / Coronary artery and umbilical vein endothelial cells) MMP-2 by HCAEC cells

6. Conclusions: DADS is able to inhibit the production of several MMPs particularly in HCAEC cells AM is less potent than DADS Garlic powder (PRI-200, MOR-200) has a similar effect Garlic powder (PRI-0) is less effective Garlic preparations might influence the MMP-TIMP balance Questions: What is the effect of garlic material on the production of TIMPs ?

7. Effect of garlic material on plasma lipids in transgenic mice Garlic powder (Li111) and DADS significantly lowered food intake and body weight gain Neither garlic powders (Li; MOR200) nor DADS or Allicin significantly changed serum cholesterol and TGL levels, blood pressure or platelet activation. (TNO the Netherlands / additions to different diets) Results:

8. Metabolism in vitro of DADS S S DADS,diallyldisulfide S S O DADSO,allicin SH AM, Allyl mercaptan S S-glutathion AGS,Allylglutathionylsulfure AMSO 2,Allylmethylsulfone S O O Allyl methyl sulfur AMSO, allyl methyl sulfoxide O S S (INRA Dijon / rat liver microomes, cytosol)

9. DADS: short half life time many metabolites  Which is the real active molecule in vivo ?

10. Metabolites in plasma 1- Early 2- long time - Very low levels of AM and AMS - Rapid appearance of AMSO and AMSO 2 (10 min.) - Peaks at the second day

12. Comparison of commercial garlic preparations and dissolution studies There are considerable differences in alliin and g- glutamylpeptides in garlic preparations. Dissolution around pH 8 results in rapid conversion of alliin, but only 60-70% is found as allicin. G- glutamylpeptides dissolve rapidly and remain essentially stable for 2 h. (Lichtwer Berlin / HPLC analysis) Results:

13. Modulation of Cholesterol Biosynthesis by Garlic Compounds Univ. of Leipzig / in vitro study in hepatocytes 0 100 200 Stim Inhib Sulfate fertilization

14. Stimulation and inhibition of cholesterol biosynthesis in Hepatocytes and HepG2 Cells by garlic powder extracted subsequently with different solvents

15. Separation scheme for garlic compounds with different effects on hepatic cholesterol biosynthesis: Garlic or Garlic Powder Solvent 2 /evapuration / reuptake Solvent 1 Fraction with very strong stimulating effect 1.Pellet Solvent 1 2.Pellet Fraction with strong inhibiting effect Needs to be kept under nitrogen

16. Conclusions: Garlic contains (two) compounds with opposite effects on cholesterol biosynthesis Questions: What is the chemical nature of the responsible compounds? Are tthese compounds effective in vivo? Do these compounds have effects on other cellular functions than cholesterol biosynthesis?

17. GST DADS AGS allyl glutathionyl sulfide AM allyl mercaptan AMS allyl methyl sulfide CYP 2B1/2 CYP 2E1 FMO AMSO2 allyl methyl sulfone AMSO allyl methyl sulfoxide DADSO Compounds obtained in presence of subcellular fractions Compounds obtained in an isolated perfused liver MT