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We have previously determined that sustained phospholipase D (PLD) activation is associated with differentiation induction in primary mouse epidermal keratinocytes. We therefore investigated the effect of two bacterial PLD on keratinocyte proliferation and differentiation. We found that Streptomyces sp. PLD was much less potent at inhibiting proliferation than S. chromofuscus PLD, with a half-maximal inhibitory concentration of 0.05 versus less than 0.001 IU per ml for S. chromofuscus PLD. Similarly, S. chromofuscus PLD stimulated transglutaminase activity more effectively and potently than S. sp. PLD. When we examined the formation of products by the two PLD, we found that the S. sp. PLD showed higher activity at all concentrations. Whereas the PLD from S. sp. is relatively inactive on sphingomyelin, S. chromofuscus PLD is known to hydrolyze both glycerophospholipids and sphingomyelin. Based on recent data indicating a role for ceramide in regulating cell growth and differentiation, we hypothesized that the ability of S. chromofuscus PLD to hydrolyze sphingomyelin might underlie its greater potency. Therefore, we examined the effect of exogenous sphingomyelinase and synthetic ceramides on DNA synthesis. We found that sphingomyelinase exhibited a potent concentration-dependent effect on [3H]thymidine incorporation, much like S. chromofuscus PLD. Synthetic cell-permeable ceramides (C6- and C2-ceramide) also concentration dependently inhibited DNA synthesis, with a half-maximal inhibitory concentration of approximately 12 microM. Finally, we obtained evidence suggesting that ceramide is generated in response to a physiologically relevant agent, because tumor necrosis factor-alpha, a known effector of sphingomyelin turnover in other systems and a cytokine that is produced and released by keratinocytes, increased ceramide levels in primary epidermal keratinocytes.

A filiform stenosis of the popliteal artery was examined by sonography and the reason for the cystic lesion in the arterial wall was found to be an uncommon type of cystic degeneration of the adventitia. The narrow lumen could be demonstrated by sonography and the lesion was punctured under ultrasound control. A viscous secretion was removed and the patient's claudication was cured without recourse to surgery.

The Problem of an Adequate Surgical Approach: Frequency of malignant teratomas is, according to the literature, 2%-10%. Examining 194 own cases (1983-1993) it was 1.5%. We found one squamous cell carcinoma (0.5%). Additionally we found 2 immature teratomas (1%). We point out the different biological behaviour of malignant mature teratomas and immature teratomas. We agree with the majority of authors that the method of choice is the intact removal of all teratomas without iatrogen rupture or contamination of the abdominal cavity by contents of the teratoma. This adequate surgical procedure can and should be performed by laparotomy or laparoscopy with endobag. The often practised method of cutting open the cyst during laparoscopy, sucking off the contents or cutting the teratoma into pieces, has been proven to lead to implantation and worsening the prognosis in case of a malignant teratoma. Even the rinsing of the abdominal cavity, usually carried out with this method, could not compensate always for the disadvantage of this "dirty" endoscopical method compared with usual oncological standards. This is pointed out by case reports in the literature and the first analysis of a German survey with early-follow-up of 192 laparoscopically managed ovarian malignancies [11a]. The principle of intact removal of every teratoma should again be kept in mind.

For the first time, a weak clinical efficacy of a 12-week therapy with garlic powder (daily dose, 800 mg) is demonstrated in patients with peripheral arterial occlusive disease stage II. The increase in walking distance in the verum group by 46 m (from 161.0 +/- 65.1 to 207.1 +/- 85.0 m) was significantly higher (P < 0.05) than in the placebo group (by 31 m, from 172.0 +/- 60.9 to 203.1 +/- 72.8). Both groups received physical therapy twice a week. The diastolic blood pressure, spontaneous thrombocyte aggregation, plasma viscosity, and cholesterol concentration also decreased significantly. Body weight was maintained. It is quite interesting that the garlic-specific increase in walking distance did not appear to occur until the 5th week of treatment, connected with a simultaneous decrease in spontaneous thrombocyte aggregation. Therefore, garlic may be an appropriate agent especially for the long-term treatment of an incipient intermittent claudication.

A platelet-inhibiting effect is described for garlic. In this double-blind, placebo-controlled study on 60 voluntary subjects with cerebrovascular risk factors and constantly increased platelet aggregation it was demonstrated that the daily ingestion of 800 mg of powdered garlic (in the form of coated tablets) over 4 weeks led to a significant inhibition of the pathologically increased ratio of circulating platelet aggregates and of spontaneous platelet aggregation. The ratio of circulating platelet aggregates decreased by 10.3%, from 1.17 +/- 0.08 to 1.05 +/- 0.11 (P < 0.01), and spontaneous platelet aggregation by 56.3%, from 40.7 +/- 23.3 to 17.8 +/- 23.2 degrees (P < 0.01) during the garlic phase. There were no significant changes in the placebo group. The parallel group comparison (garlic versus placebo) revealed a significantly different ratio of circulating platelet aggregates after 4 weeks of treatment (P < 0.05). After the 4-week wash-out phase the values increased again to 1.19 +/- 0.32 and 34.9 +/- 28.7 degrees, reaching the initial values (run-in phase prior to the ingestion of garlic). Since garlic is well tolerated it would be worth testing it in a controlled clinical trial for usefulness in preventing disease manifestations associated with platelet aggregation.

In a randomized placebo-controlled double-blind cross-over study it could be shown that 5 h after the administration of garlic powder (Kwai, Sapec; total dose of 900 mg garlic powder) a significant increase in capillary skin perfusion by 55% occurs in the healthy volunteers. Placebo did not cause any changes. The difference between the two study phases is also significant. The increased erythrocyte velocity results from vasodilation of precapillary arterioles which increases diameter of erythrocyte column by an average of 8.6%. Simultaneously inflow of interstitial fluidity accompanied by a significant decrease in haematocrit and plasma viscosity occurs (rheoregulation).

The DNA encoding protease Do was isolated from an E. coli genomic DNA library in lambda gt11, and cloned into a Bluescript plasmid. The cells transformed with the recombinant plasmid were able to overproduce protease Do and grew normally. A mutant lacking the protease activity was also isolated by interrupting the chromosomal DNA with the kan gene. The mutant showed a prolonged lag period and reduced ability to degrade cell proteins as compared to its wild type. Moreover, they were unable to survive at high temperatures, similarly to the htrA mutants. These results suggest that protease Do may play an important role in the intracellular protein breakdown and is essential for survival at high temperatures. Identity of protease Do with the htrA gene product is discussed.

Significant positive effects could be achieved in a placebo-controlled double-blind study through the administration of 800 mg of garlic powder over a period of four weeks. Spontaneous thrombocyte aggregation disappeared, the microcirculation of the skin increased by 47.6% (from 0.63 +/- 0.13 to 0.93 +/- 0.22 mm/s), plasma viscosity decreased by 3.2% (from 1.25 +/- 0.34 to 1.21 +/- 0.43 mPas), diastolic blood pressure by 9.5% (from 74 +/- 9 to 67 +/- 5 mmHg), and blood glucose concentration by 11.6% (from 89.4 +/- 8.8 to 79.0 +/- 11.9 mg/dl). The vascular protection of garlic as atherosclerosis prevention by influencing the mentioned risk parameters for cardiovascular diseases must be pointed out. Especially interesting is the thrombocyte aggregation inhibiting effect. Thus, the application of garlic may be useful in case of acetylsalicyclic acid intolerance.

Induction of protease La was found to increase to higher extent in E. coli that had been treated with canavanine for longer period. However, hydrolysis of canavanine-containing proteins occurred rapidly but at nearly an identical rate regardless of the period of canavanine-treatment. Exposure of E. coli to heat also raised the level of protease La but showed little effect on overall rate of proteolysis. These results suggest that induction of protease La under stress occurs as a part of heat shock response but not necessarily for elimination of denatured or abnormal proteins.