The efficacy and safety of DW1601 in patients with acute bronchitis: a multi-center, randomized, double-blind, phase III clinical trial.

BACKGROUND/AIMS: DW1601, an oral fixed dose combination syrup composed of DW16011 and Pelargonium sidoides, was developed to enhance the symptom relief effect in patients with acute bronchitis. We evaluated the efficacy and safety of DW1601 compared to DW16011 or P. sidoides for treatment of acute bronchitis using a randomized, double-blind, placebocontrolled, multi-centre trial design. METHODS: A total of 204 patients with acute bronchitis was randomized 1:1:1 to receive DW1601 (n = 67), DW16011 (n = 70), or P. sidoides (n = 64) for 7 days. The primary outcome was efficacy of DW1601 compared to DW16011 or P. sidoides in reducing the total bronchitis severity score (BSS) at day 4 of treatment. Secondary endpoints were changes in total and symptomspecific BSS, response rate and patient satisfaction rate. Safety analysis was assessed at day 7. RESULTS: At 4 days after medication, decrease of total BSS from baseline was significantly greater in the DW1601 group than in the DW16011 group (-3.51 ± 0.18 vs. -2.65 ± 0.18, p = 0.001) or P. sidoides group (-3.56 ± 0.18 vs. -2.64 ± 0.19, p < 0.001). In addition, the BSS total score at day 7 and the BSS cough and sputum component scores at days 4 and 7 were significantly more improved with DW1601 treatment compared with the DW16011 group or P. sidoides group. Participants treated with DW1601 showed higher rates of response and satisfaction than control groups (response rate, DW1601, 100% vs. DW16011, 85.7% vs. P. sidoides, 85.9%; satisfaction rate, DW1601, 92.6% vs. DW16011, 82.9% vs. P. sidoides, 81.2%). Significant adverse events were not observed in the DW1601 group. CONCLUSION: DW1601 is superior to DW16011 or P. sidoides in improving symptoms of acute bronchitis.

Effects of a Mixture of Ivy Leaf Extract and Coptidis rhizome on Patients with Chronic Bronchitis and Bronchiectasis.

BACKGROUND: Hederacoside C from ivy leaf dry extracts (HH) and berberine from Coptidis rhizome dry extracts (CR) can be combined (HHCR) as a herbal product. Previous studies have demonstrated that HHCR has antitussive and expectorant effects in animal models of respiratory disease. However, the therapeutic effects of HHCR on respiratory diseases in humans have not been well-studied. Therefore, we aimed to clarify the effectiveness of HHCR in patients with chronic bronchitis and bronchiectasis. METHODS: This was a multicenter (10 university teaching hospitals), open-label, prospective, single-arm, observational study. Consecutive patients with chronic bronchitis and bronchiectasis were included. Patients were orally treated with HHCR daily for 12 weeks. St. George's Respiratory Questionnaire (SGRQ) scores and bronchitis severity scores (BSS) were measured at baseline and at the end of the 12-week study. RESULTS: In total, 376 patients were enrolled, of which 304 were finally included in the study, including 236 males and 68 females with a median age of 69 years (range: 37-88 years). After 12 weeks of HHCR treatment, there was a significant improvement in SGRQ score (baseline, 32.52 ± 16.93 vs. end of study, 29.08 ± 15.16; p < 0.0001) and a significant reduction in BSS (baseline, 7.16 ± 2.63 vs. end of study, 4.72 ± 2.45; p < 0.0001). During the study, 14 patients concomitantly used an inhaled corticosteroid and 83 patients used an inhaled bronchodilator. HHCR also had significant positive effects on these patients in terms of SGRQ score and BSS. No serious adverse drug reactions occurred during HHCR treatment. CONCLUSIONS: treatment with HHCR improved the SGRQ score and BSS in patients with chronic bronchitis and bronchiectasis. HHCR may be a new therapeutic option for chronic bronchitis and bronchiectasis. Large-scale, randomized, double-blind, placebo-controlled clinical trials are warranted.

Preparation of High Purity Δ5-Olefinic Acids from Pine Nut Oil via Repeated Lipase-Catalyzed Esterification.

Δ5-Olefinic acids have been characterized in gymnosperm plants and have been reported to have several biological health benefits. Δ5-Olefinic acids from pine nut oil were effectively concentrated by repeated lipase-catalyzed esterification. The pine nut oil contained three major Δ5-olefinic acids, namely taxoleic acid (C18:2 Δ5,9), pinolenic acid (C18:3 Δ5,9,12), and sciadonic acid (C20:3 Δ5,11,14). The fatty acids present in pine nut oil were selectively esterified with ethanol using Lipozyme RM IM from Rhizomucor miehei as a biocatalyst. The Δ5-olefinic acids were concentrated in the unesterified fatty acid fraction. The optimum molar ratio of the substrates (fatty acid:ethanol), temperature, the enzyme loading, and the reaction time were 1:7, 25°C, 5% of total substrate weight, and 6 h, respectively. There was no significant effect in the concentration of Δ5-olefinic acids when water was added in the reaction mixture. The same protocol and optimum conditions were employed for two times repeated lipase-catalyzed esterifications. In first lipase-catalyzed esterification, the Δ5-olefinic acids content in the pine nut oil increased from 17 mol% to 51 mol% with a yield of 40 mol%. In a second lipase-catalyzed esterification, with the Δ5-olefinic acids-concentrated fatty acids obtained from the first reaction as the substrate, the Δ5-olefinic acids content increased to 86 mol% with a yield of 15 mol%. Finally, a maximum Δ5-olefinic acids content of ca. 96 mol% with a yield of 6 mol% was obtained via a third lipase-catalyzed esterification.

Optimization of culture conditions for production of a novel cold-active lipase from Pichia lynferdii NRRL Y-7723.

Lipases with abnormal properties such as thermostability, alkalinity, acidity, and cold activity receive industrial attention because of their usability under restricted reaction conditions. Most microbial cold-active lipases originate from psychrotrophic and psychrophilic microorganisms found in Antarctic regions, which has led to difficulties in the practical production of cold-active lipase. Recently, a mesophilic yeast, Pichia lynferdii NRRL Y-7723, was reported to produce a novel cold-active lipase. This study focused on optimization of environmental factors, while giving particular attention to the relationships between given factors and incubation time, to maximize the production of a novel cold-active lipase from P. lynferdii NRRL Y-7723. Maximum lipase production was highly dependent on the incubation time at a given environmental factor. Lipase production varied with incubation time at a given temperature, and 20 °C was selected as the optimum temperature for lipase production. Fructose was selected as the best carbon source, and maximum lipase production was obtained when it was present at 0.7% (w/v). Yeast extract was an efficient organic nitrogen source, with maximum lipase production occurring at 0.9% (w/v). Specifically, at the optimum yeast extract level the lipase production was >10 times higher than the productivity under standard conditions. All natural oils tested showed lipase production, but their maximum productivities varied according to incubation time and oil species.

Production of 7,10-dihydroxy-8(E)-octadecenoic acid from olive oil by Pseudomonas aeruginosa PR3.

Microbial modification of naturally occurring materials is one of the efficient ways to add new values to them. Hydroxylation of free unsaturated fatty acids by microorganism is a good example of those modifications. Among microbial strains studied for that purpose, a new bacterial isolate Pseudomonas aeruginosa PR3 has been well studied to produce several hydroxy fatty acids from different unsaturated fatty acids. Of those hydroxy fatty acids, 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was efficiently produced from oleic acid by strain PR3. However, it was highly plausible to use vegetable oil containing oleic acid rather than free oleic acid as a substrate for DOD production by strain PR3. In this study, we firstly tried to use olive oil containing high content of oleic acid as a substrate for DOD production. DOD production from olive oil was confirmed by structural determination with GC, TLC, and GC/MS analysis. DOD production yield from olive oil was 53.5%. Several important environmental factors were also tested. Galactose and glutamine were optimal carbon and nitrogen sources, and magnesium ion was critically required for DOD production from olive oil. Results from this study demonstrated that natural vegetable oils containing oleic acid could be used as efficient substrate for the production of DOD by strain PR3.

Effect of surfactant on the production of oxygenated unsaturated fatty acids by Bacillus megaterium ALA2.

Bacillus megaterium ALA2 NRRL B-21660 has been well studied for the production of many oxygenated unsaturated fatty acids from linoleic acid. Its major product, 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17-THOA), inhibited the growth of many plant pathogenic fungi. However, we have been unable, until now, to demonstrate 12,13,17-THOA production in a fermentor. Here, we have investigated the effect of surfactants on 12,13,17-THOA production. Surfactant types (SO-25, Tween-80 and Triton X-100) at various concentrations were evaluated for their effects on cell growth and production of 12,13,17-THOA. Triton X-100 decreased cell growth and 12,13,17-THOA production while 1% Tween-80 increased 12,13,17-THOA production more than twofold over control. In a pH and dissolved oxygen (DO) controlled fermentor and with 1.0% Tween-80, both 12,13,17-THOA production and productivity increased to 553 mg/L and 24.0mg/L/hour, respectively. We further conducted THOA production in a fermentor without the control of pH and DO, but with aeration through medium surface rather than by Spurger, and with various amounts of Tween-80 to compare with the results obtained in flask runs. Both additions with 0.1% and 0.5% Tween-80 and the control runs produced poor amounts of product 12,13,17-THOA. The maximum 12,13,17-THOA production was observed at 1.0% Tween-80 at a yield of 650 mg/L. Thus fermentor production of 12,13,17-THOA was successfully demonstrated.

Lipase-catalyzed acidolysis of olive oil with capric acid: effect of water activity on incorporation and acyl migration.

Structured lipids were synthesized by acidolysis of olive oil and capric acid with an immobilized lipase (Lipozyme TL IM) from Thermomyces lanuginosus. The acidolysis reaction was carried out by incubating a 1:3 molar ratio of olive oil and capric acid under solvent-free reaction systems at 50 degrees C. The effect of water activity on the incorporation of capric acid was investigated, and the tested water activity range was between 0.22 and 0.80. Capric acid incorporation into triacylglycerols of the olive oil increased as the water activity increased, but the degree of acyl migration also increased. Also, the degree of acyl migration of modified olive oils with a similar degree of incorporation was investigated. High degrees of acyl migration occurred at water activities of 0.22 and 0.32 for the degree of incorporation of ca. 50 mol %. Only 8 h of reaction time was required to achieve incorporation of ca. 50 mol % at a water activity of 0.80, and the lowest acyl migration occurred at the same water activity. These results suggest that acyl migration can be efficiently minimized by a shorter reaction time at higher water activity.

Green tea extract inhibits paraquat-induced pulmonary fibrosis by suppression of oxidative stress and endothelin-l expression.

Paraquat-induced pulmonary fibrosis involves two factors, direct injury by oxygen free radicals and indirect injury by inflammatory cells and fibroblasts. Endothelin-1 (ET-1) has been shown to act as a mediator of pulmonary fibrosis, and its formation increases during oxidative stress. We investigated whether green tea extract (GTE), which has antioxidant properties, inhibits paraquat-induced pulmonary fibrosis and whether ET-1 is involved in this process. Paraquat (0.3 mg/kg) was instilled into the right lungs of rats, following which the rats were either not further treated (Group P, n = 7), or they were administered 1% GTE mixed with feed (Group PG; n = 7) or the ET(A) receptor antagonist ZD2574 (10 mg/kg through gavage; Group PZ; n = 7) for two weeks. As control, we used rats instilled with saline (Group N; n = 6). Two weeks after paraquat instillation, we assayed the degree of pulmonary fibrosis by light microscopic morphometry and hydroxyproline content; lipid peroxidation as a marker of oxidative stresses by measurement of malondialdehyde (MDA); ET-1 by immunohistochemistry; and prepro-ET-1 mRNA expression by reverse transcription-polymerase chain reaction. Compared with Group N, significant pulmonary fibrosis was observed in Group P, accompanied by increases in MDA, ET-1, and prepro-ET-1 mRNA expression. Compared with Group P, Group PG showed significant decreases in pulmonary fibrosis, along with decreases in MDA, ET-1, and prepro-ET-1 mRNA expression. We also observed significant decreases in pulmonary fibrosis in Group PZ compared with Group P. These findings suggest that GTE inhibits paraquat-induced pulmonary fibrosis by suppression of oxidative stress and ET-1 expression.

Antioxidant property of an ethanol extract of the stem of Opuntia ficus-indica var. saboten.

An ethanol extract of the stem of Opuntia ficus-indica var. saboten (OFS) was assessed to determine the mechanism(s) of its antioxidant activity. The ethanol extract exhibited a concentration-dependent inhibition of linoleic acid oxidation in a thiocyanate assay system. In addition, the OFS extract showed dose-dependent free-radical scavenging activity, including DPPH radicals, superoxide anions (O(2)(*-)), and hydroxyl radicals (*OH), using different assay systems. The OFS ethanol extract was also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in a Fenton's reaction mixture. Furthermore, the extract showed significant (p < 0.01) dose-dependent protection of mouse splenocytes against glucose oxidase-mediated cytotoxicity. Finally, the OFS extract was characterized as containing a high amount of phenolics (180.3 mg/g), which might be the active compounds responsible for the antioxidant properties of the OFS extract.