Immunomodulatory effects of aged garlic extract.

Using various kinds of models, we examined the effects of aged garlic extract (AGE) on immune functions. In the immunoglobulin (Ig)E-mediated allergic mouse model, AGE significantly decreased the antigen-specific ear swelling induced by picryl chloride ointment to the ear and intravenous administration of antitrinitrophenyl antibody. In the transplanted carcinoma cell model, AGE significantly inhibited the growth of Sarcoma-180 (allogenic) and LL/2 lung carcinoma (syngenic) cells transplanted into mice. Concomitantly, increases in natural killer (NK) and killer activities of spleen cells were observed in Sarcoma-180--bearing mice administered AGE. In the psychological stress model, AGE significantly prevented the decrease in spleen weight and restored the reduction of anti-SRBC hemolytic plaque-forming cells caused by the electrical stress. These studies strongly suggest that AGE could be a promising candidate as an immune modifier, which maintains the homeostasis of immune functions; further studies are warranted to determine when it is most beneficial.

Pharmacologic activities of aged garlic extract in comparison with other garlic preparations.

We investigated the pharmacologic activities of four garlic preparations, raw garlic juice (RGJ), heated garlic juice (HGJ), dehydrated garlic powder (DGP) and aged garlic extract (AGE). The study used three animal models, i.e., testicular hypogonadism (hypospermatogensis and impotence) induced by warm water treatment, intoxication of acetaldehyde and growth of inoculated tumor cells. RGJ was found to be effective only in recovery of testicular function. The efficacy of HGJ was observed in three models; however, it did not improve impotence. DGP was effective in recovery of spermatogenesis and stimulated acetaldehyde detoxification. Significant beneficial effects of AGE were found in all three models. Although all four garlic preparations significantly enhanced natural killer (NK) and killer cell activities of the spleen cells of tumor-bearing mice, only AGE and HGJ inhibited the growth of inoculated tumor cells. These results suggest that different types of garlic preparations have different pharmacologic properties, and among the four garlic preparations studied, AGE could be the most useful garlic preparation.

Prevention of psychological stress-induced immune suppression by aged garlic extract.

We determined the effect of Aged Garlic Extract (AGE) on damage caused to immune function by a psychological stress using a communication box. After four days of a psychological stress, a decrease in spleen weight and spleen cells was observed in the psychological stress-exposed mice as compared normal mice (non-stress). AGE significantly prevented the decreases in spleen weight and cells. Additionally, AGE significantly prevented the reduction of hemolytic plaque-forming-cells in spleen cells and anti-SRBC antibody titer in serum caused by this psychological stress. Moreover, a reduction in NK activities was observed in the psychological stress-exposed mice as compared with normal mice (non-stress), whereas NK activities in the AGE administered mice were almost the same as normal mice (non-stress). These results indicate that psychological stress qualitatively and quantitatively impairs immune function, and that AGE is extremely useful for preventing psychologically-induced damage.

Immunomodulation and antitumor activities of Aged Garlic Extract.

We found that Aged Garlic Extract (AGE) could be a significant immuno-potentiator, and could exhibit anti-tumor activities through immune modulation. Consequently, AGE stimulated the proliferation of mouse spleen cells and the release of cytokines, such as IL-2, TNF-alpha and IFN-gamma, increased NK activities, and enhanced phagocytosis of peritoneal macrophages. AGE treatment also stimulated the reactivity of lymphocytes in response to cytokines or mitogens. AGE was far superior to PSK in IL-2 induction, but slightly inferior to PSK in nitric oxide induction. AGE, as effectively as PSK (Krestin), significantly inhibited the growth of Sarcoma-180 (allogenic) and LL/2 lung carcinoma (syngenic) cells transplanted into mice. Concomitantly, increases in NK and killer activities of spleen cells were observed in Sarcoma-180 bearing mice treated with AGE. These results strongly suggest that AGE is as effective as PSK, and could serve as a potent biological response modifier on NK cells and T lymphocytes, and subsequently inhibit the growth of transplanted tumors.

Anti-allergic effects of aged garlic extract.

To examine the effect of Aged Garlic Extract (AGE) on the function of mast cells and activated T lymphocytes, we adopted the in vitro histamine release system, the in vivo IgE mediated skin reaction system and the in vivo late phase reaction system. Consequently, at 1.25, 2.5, and 5.0% (v/v), AGE dose-dependantly inhibited the antigen specific histamine release by mouse anti-TNP monoclonal antibody and TNP-BSA hapten carrier complex against rat basophil cell line RBL-2H3 by 50, 80, and 90 percent, respectively. In the IgE mediated skin reaction system, repeated or single intragastric administration of AGE (10 ml/kg), decreased by 25-45% the antigen specific ear swelling which was induced by a picryl chloride ointment applied to the ear of mice also given an intravenous administration of anti-TNP antibody IgE ascites. In the late phase reaction system, repeated or single intragastric administration of AGE (10 ml/kg) suppressed by 45-55% the antigen specific ear swelling induced by a secondary challenge to the ear of mice given a picryl chloride ointment seven days prior. These results suggest that AGE application could modify, directly or indirectly, the function of mast cells, basophils and activated T lymphocytes which play a leading role in allergic cascade reactions including inflammation.

Tautomycin: an inhibitor of protein phosphatases 1 and 2A but not a tumor promoter on mouse skin and in rat glandular stomach.

Tautomycin isolated from Streptomyces spiroverticillatus is an inhibitor of protein phosphatases 1 and 2A. Tautomycin induced hyperphosphorylation of cytokeratin peptides in human keratinocytes (PHK 16-I cells) 30 times less strongly than did okadaic acid. Repeated applications of tautomycin (30 micrograms, 40 nmol/application) did not induce tumor promotion in a two-stage carcinogenesis experiment on mouse skin initiated with 7,12-dimethylbenz[a]anthracene, whereas okadaic acid (1 microgram, 1.2 nmol/application) as a control induced tumor promotion strongly. As for mucosa of rat glandular stomach, tautomycin induced ornithine decarboxylase 4 h after intubation into the stomach. The tumor-promoting activity of tautomycin was next studied in the glandular stomach initiated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Administration of tautomycin in the diet (1 mg rat-1 day-1), from week 9 to week 52 of the experiment, inhibited rather than enhanced tumor development in the glandular stomach initiated with MNNG. The percentages of tumor-bearing rats of the groups treated with MNNG plus tautomycin, MNNG alone, and tautomycin alone were 20.0%, 40.6%, and 0% respectively in week 52. The reason for the absence of tumor-promoting activity of tautomycin was studied in relation to tumor necrosis factor alpha (TNF alpha), an endogenous tumor promoter. We found that tautomycin neither enhanced TNF alpha mRNA expression in mouse skin nor induced TNF alpha release in a human stomach cancer cell line (KATO III cells), whereas okadaic acid did both. These results indicate that not all inhibitors of protein phosphatases are tumor promoters, and suggest that tumor promotion of the okadaic acid class of compounds is mediated by TNF alpha.

Clinicopathological correlations of visual depth perception in patients with cerebrovascular disease.

Visual depth perception in 100 patients with cerebrovascular disease was evaluated using the Titumus stereotest. Twelve patients showed depth perception impairment. CT scans revealed that the lesion was located on the right hemisphere in 6 patients and on the left hemisphere in 3 patients, and that the remaining 3 patients had multiple infarctions. 123I SPECT was performed in 7 patients with moderate to severe depth perception impairment, of whom 6 patients showed a reduced blood flow in the posterior half of both the right and left cerebral hemispheres. Moderate to severe impairment of depth perception was more frequently observed in patients with a lesion in the right hemisphere or in the posterior half of either hemisphere.

Apparent "activation" of protein kinases by okadaic acid class tumor promoters.

A cytosolic fraction of mouse brain gave two peaks of protein kinase activity on DEAE-cellulose column chromatography. The first peak of protein kinase corresponded to protein kinase C. The second peak contained protein kinases that were "activated" dose-dependently by the okadaic acid class tumor promoters, okadaic acid and dinophysistoxin-1. This "activation" was not achieved by other tumor promoters, such as 12-0-tetradecanoyl-phorbol-13-acetate, teleocidin, aplysiatoxin, or palytoxin. In addition, the second peak contained phosphatases. The phosphate liberation from phosphorylated histone type III-S by incubation with the second peak was inhibited by okadaic acid or dinophysistoxin-1, dose-dependently. The resulting apparent "activation" of protein kinases by okadaic acid is indicated and would imply a new pathway of tumor promotion on mouse skin.

Diarrhetic shellfish toxin, dinophysistoxin-1, is a potent tumor promoter on mouse skin.

Dinophysistoxin-1, 35-methylokadaic acid, is a causative agent of diarrhetic shellfish poisoning. The biological activities and tumor-promoting activity of dinophysistoxin-1 were studied together with those of okadaic acid and 7-O-palmitoyl okadaic acid. Dinophysistoxin-1 is a skin irritant and induces ornithine decarboxylase in mouse skin with the same potency as okadaic acid. 7-O-Palmitoyl okadaic acid induced a lower activity than the other compounds. Dinophysistoxin-1 inhibited the specific [3H]okadaic acid binding to a particulate fraction of mouse epidermis. The binding affinities of dinophysistoxin-1 and okadaic acid to a particulate fraction were almost the same. Dinophysistoxin-1 showed a tumor-promoting activity as strong as that of okadaic acid in a two-stage carcinogenesis experiment on mouse skin. The percentages of tumor-bearing mice in the groups treated with 100 micrograms of 7,12-dimethylbenz[a]anthracene (DMBA) followed by 5 micrograms of dinophysistoxin-1, twice a week, and with DMBA followed by 5 micrograms of okadaic acid twice a week were 86.7% and 80.0% in week 30, respectively. The average number of tumors per mouse was 4.6 in the former group and 3.9 in the latter. Dinophysistoxin-1 and okadaic acid act on cells through different pathways from the 12-O-tetradecanoylphorbol-13-acetate-type tumor promoters.

S-(1,2-dicarboxyethyl)glutathione and S-(1,2-dicarboxyethyl)L-cysteine in lens.

S-(1,2-Dicarboxyethyl)glutathione and S-(1,2-dicarboxyethyl)L-cysteine were determined by high performance liquid chromatography after reaction with 2,4-dinitrofluorobenzene. By this method the former could be determined in the range 4.05 mumol/l-815 mumol/l, and the latter in the range 1.45 mumol/l-1.45 mmol/l. The recovery from cattle lens homogenate was 90.0 +/- 3.2% for S-(1,2-dicarboxyethyl)glutathione and 95.3 +/- 3.1% for S-(1,2-dicarboxyethyl)L-cysteine. Using this method S-(1,2-dicarboxyethyl)-glutathione and S-(1,2-dicarboxyethyl)L-cysteine were determined in lenses of several vertebrates and in rat lens during cataract formation by galactose.