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Indications for post-translational regulation of Vitis vinifera L. arginine decarboxylase.

In order to study arginine decarboxylase regulation, we produced an antiserum against a hybrid of a 615 amino acid residue fragment of grapevine arginine decarboxylase cDNA with maltose-binding protein. The antiserum generated recognized mainly a protein band of ca. 80 kDa in extracts from grapevine tissues. Extracts from leaves and internodes in different developmental stages showed differences in the quantity of the 80 kDa band recognized by the antiserum. However, these differences did not correspond with changes in arginine decarboxylase specific activity. Furthermore, western blot analysis of extracts from cell cultures, where enzyme-specific activity was induced or repressed, did not reveal respective changes in the quantity of the 80 kDa protein band. Digestion of the hybrid by the specific protease factor Xa resulted in a polypeptide of 90 kDa instead of the expected two polypeptides of 43 and 66 kDa. Finally, western blot analysis of shoot extract incubated with factor Xa or the hybrid protein previously digested by factor Xa revealed that factor Xa-digested hybrid protein cleaved the 80 kDa band, resulting in two bands of ca. 38 and 40 kDa, whereas factor Xa alone did not affect it. These results suggest that arginine decarboxylase protein levels and/or activity is post-translationally regulated, as has been shown for other enzymes of polyamine biosynthesis.

Animals↗

Efficacy of orally administered extract of red vine leaf AS 195 (folia vitis viniferae) in chronic venous insufficiency (stages I-II). A randomized, double-blind, placebo-controlled trial.

UNLABELLED: Red vine leaf extract (RVLE) is a herbal medicine containing several flavonoids, with quercetin-3-O-beta-glucuronide and isoquercitrin (quercetin-3-O-beta-glycoside) as the main components. OBJECTIVE: To assess the efficacy and safety of once-daily doses of 360 and 720 mg RVLE (pharmaceutical extract code AS 195; Antistax Venenkapseln) compared to placebo in patients with stage I and incipient stage II chronic venous insufficiency (CVI). DESIGN: A 12-week, randomized, double-blind, placebo-controlled, parallel-group, multi-center study. PATIENTS: Male and female outpatients aged 25 to 75 years with stage I to stage II CVI (i.e. without extensive trophic changes), not having any other significant medical conditions and not treated with compression stockings, diuretics or other drugs affecting fluid balance. INTERVENTION: Patients were randomly assigned to a double-blind treatment with placebo, 360 mg AS 195 or 720 mg AS 195 once daily for 12 weeks, preceded and followed by a single-blind 2-week placebo treatment for baseline run-in and end-of-trial washout, respectively. Study criteria were evaluated at baseline, after 6 and 12 weeks of treatment and 2 weeks after discontinuation of treatment. RESULTS: Of the 260 patients enrolled and randomized, 219 completed the study in accordance with the protocol. In the intention-to-treat analysis (N = 257), the mean (+/- SD) lower leg volume (measured by water displacement plethysmography) of the patients treated with placebo (N = 87) increased by 15.2 +/- 90.1 g (displaced water mass) and by 33.7 +/- 96.1 g after 6 and 12 weeks compared to baseline. In contrast, for patients treated with AS 195, lower leg volume decreased, and after 12 weeks of treatment, the difference in mean lower leg volume between the active treatment groups and the placebo group was -75.9 g (95% CI: -106.1 to -45.8 g) and -99.9 g (95% CI: -130.3 to -69.6 g) for the group treated with 360-mg AS 195 (N = 86) and 720-mg AS 195 (N = 84), respectively. The changes in calf circumference showed a similar pattern: in patients treated with AS 195, both the higher dose (720 mg) and, albeit to a lesser extent, the lower dose (360 mg) resulted in a clear reduction in circumference over time, whereas, circumference remained largely unchanged in patients treated with the placebo (95% CI of the estimated treatment effects vs. placebo after 12 weeks: -1.40 to -0.56 cm and -1.73 to -0.88 cm for 360 and 720 mg AS 195, respectively). These differences were statistically significant (p < 0.001). The reductions in ankle circumference were qualitatively similar but quantitatively less marked. Subjectively, there was an improvement in key CVI symptoms (VAS) at 6 weeks with all treatments, but a further improvement at week 12 was seen only in the active treatment groups; at 12 weeks, the changes compared to baseline were significantly greater (p < 0.001) in both active treatment groups than in the placebo group. The treatments were well tolerated; Adverse events were rare and usually mild. Two adverse events (AEs) during treatment with the placebo led to hospitalization and were hence labeled as 'serious'. Three further patients were withdrawn because of AEs which occurred during treatment with the placebo. CONCLUSION: Once-daily doses of 360 and 720 mg AS 195 were confirmed to be safe and effective in the treatment of mild CVI, reducing significantly lower leg edema and circumference whilst improving key CVI-related symptoms to a clinically relevant extent. The edema reduction is at least equivalent to that reported for compression stockings and/or other edema-reducing agents. The higher dose was as well tolerated as the lower dose but resulted in a slightly greater and more sustained improvement.

Adult↗

Vacuolar contents of fruit subepidermal cells from vitis species.

Enzymic treatment of mature, anthocyanin-containing grape berry sub-epidermal tissues from DeChaunac grapes released intact protoplasts. Filtration of the protoplast suspension through glass wool under mild suction resulted in the release of vacuoles. The total and individual contents of anthocyanins, flavonol glycosides, hydroxycinnamic acid esters, sugars, organic acids, and cations were determined in both tissue and vacuole preparations. A method for pH determination in intact vacuoles is reported. Based on the qualitative and quantitative anthocyanin composition, the average pH of the vacuoles was determined to be 2.7 (sd +/- 0.17). The data suggest that anthocyanins are present in fruit subepidermal tissues in a noncomplexed form.

Journal Article↗

The Agrobacterium vitis T-6b oncoprotein induces auxin-independent cell expansion in tobacco.

Among the Agrobacterium T-DNA genes, rolB, rolC, orf13, orf8, lso, 6b and several other genes encode weakly homologous proteins with remarkable effects on plant growth. The 6b oncogene induces tumors and enations. In order to study its properties we have used transgenic tobacco plants that carry a dexamethasone-inducible 6b gene, dex-T-6b. Upon induction, dex-T-6b plants develop a large array of morphological modifications, some of which involve abnormal cell expansion. In the present investigation, dex-T-6b-induced expansion was studied in intact leaves and an in vitro leaf disc system. Although T-6b and indole-3-acetic acid (IAA) both induced expansion and were non-additive, T-6b expression did not increase IAA levels, nor did it induce an IAA-responsive gene. Fusicoccin (FC) is known to stimulate expansion by increasing cell wall plasticity. T-6b- and FC-induced expansion were additive at saturating FC concentrations, indicating that T-6b does not act by a similar mechanism to FC. T-6b expression led to higher leaf osmolality values, in contrast to FC, suggesting that the T-6b gene induces expansion by increasing osmolyte concentrations. Metabolite profiling showed that glucose and fructose played a major role in this increase. We infer that T-6b disrupts the osmoregulatory controls that govern cell expansion during development and wound healing.

Bacterial Proteins↗

Diversity of the limited-host range iaaH gene of Agrobacterium vitis strain Ag162.

The indole-3-acetamide hydrolase gene (iaaH) of the limited-host range strain AG162, a biotype III strain of Agrobacterium tumefaciens has been the subject of several studies and reviews, but its primary structure has not been previously reported. In the course of our own work we found that this gene hybridizes only weakly to a nucleic acid probe corresponding to the iaaH gene from a biotype I strain of A. tumefaciens. Analysis of the primary structure of the Ag162 iaaH gene revealed that it is diverse from biotype I iaaH genes and, surprisingly, also from the iaaH genes of previously characterized biotype III Agrobacterium strains.

Amidohydrolases↗

[Clinical and capillaroscopic evaluation of chronic uncomplicated venous insufficiency with procyanidins extracted from vitis vinifera].

BACKGROUND: The pharmacological treatment of non-complicated chronic venous insufficiency is a current and well-debated topic. The introduction of new products with action on the venous system, improved knowledge on the physiopathology of venous insufficiency and the possibility provided by new analytical instruments, have given new impulse to the consolidation of the clinical value of phlebotonics in this indication. METHODS: In light of this, 24 patients with non-complicated chronic venous insufficiency were treated with oral administration of Oligomeric Proanthocyanidins (Pycnogenols-OPC) 100 mg/day. To evaluate the therapeutic efficacy of the treatment, an instrumental evaluation by optical probe capillaroscope was employed in addition to the traditional subjective clinical parameters: swelling, itching, heaviness and pain. The videocapillaroscope examination was performed at the lower third of the leg and the first toe. Edema in the capillaroscopic field, the number of observable capillaries and the capillary dilatation were the parameter chosen to evaluate the efficacy of treatment. All patients completed the study with no reports of adverse events during the period of observation. RESULTS: The results obtained show a positive clinical response (improved or absent symptoms) in over 80% of patients, with significant improvement of symptoms already evident after the first 10 days of treatment. The mechanism of action of the OPCs explains the rapid reduction of the swelling of the lower limbs and correlated with this are the other evaluable symptoms: heaviness and itching. Particularly striking results were observed for itching and pain which completely disappeared during the course of therapy in 80% and 53% of the patients respectively. Noteworthy is the good correlation between the clinical and instrumental data, with improvement in a total of 70% of patients. CONCLUSIONS: The results obtained in the course of this clinical experience, with evident improvement already during the first weeks of treatment, the absence of adverse events added to the benefit of a once-a-day administration, justify the use of OPC in the treatment of non-complicated chronic venous insufficiency.

Adult↗

Influence of UV-B irradiation on the carotenoid content of Vitis vinifera tissues.

The carotenoid content was examined in leaf and berry tissues of grapevines (cv. Cabernet Sauvignon) grown either under ambient conditions or under a polyester film to reduce UV light by 98%. Total carotenoids in leaves were less in vines grown under the UV screen. Levels of beta-carotene decreased with berry development around veraison. This effect was more pronounced in vines grown under reduced UV light. The lutein content of berries appeared to remain relatively constant with berry development, but levels were decreased under the UV screen. These observations are important for the wine industry because of the biosynthetic link between carotenoids and wine flavour and aroma compounds.

Carotenoids↗