The growth-inhibitory effects of pawpaw (Asimina triloba [L.] Dunal) roots, twigs, leaves, and fruit against human gastric (AGS) and cervical (HeLa) cancer cells and their anti-inflammatory activities.

BACKGROUND: The pawpaw tree has several beneficial effects. However, no studies have been conducted to address the mechanisms underlying the cytotoxic effects of pawpaw extracts against cancer cells, and no study has investigated the anti-inflammatory effects. Hence, in this study, the growth-inhibitory effects of pawpaw (Asimina triloba [L.] Dunal) extracts against gastric (AGS) and cervical (HeLa) cancer cells and the inhibitory effects of pawpaw extracts against inflammatory factors (NO, TNF-α, IL-6, iNOS, and COX-2) were investigated. METHODS AND RESULTS: The viability of AGS and HeLa cells, the analysis of cell cycle, and the expression of apoptosis marker protein were determined using MTT assay, FACS, western blotting, and TUNEL assays. The inflammatory factors were determined using Griess method, ELISA assay kit, and RAW 264.7 cells. The IC50 values of twig and unripe fruit extracts for AGS cells were 82.01 and 100.61 µg/mL, respectively. For HeLa cells, pawpaw twig extracts exhibited the strongest ability to inhibit cervical cancer cell growth (IC50 = 97.73 µg/mL). Analysis of the cell cycle phase distribution and expression of the apoptosis regulatory proteins BCL-2, BAX, caspase-3, and PARP showed that pawpaw twig, root, and unripe fruit extracts induced Sub G1 cell cycle arrest and apoptosis of AGS and HeLa cells. In addition, the twig, root, and unripe fruit extracts of pawpaw effectively inhibited the inflammatory makers NO, TNF-α, IL-6, and iNOS. Particularly, the twig, root, and unripe fruit extracts at concentrations of 50 µg/mL exhibited > 50% inhibition of TNF-α. CONCLUSIONS: These findings indicate that pawpaw extracts are natural therapeutic agents that may be used for the prevention and treatment of gastric and cervical cancers, and encourage further studies on the anti-inflammatory potential of the pawpaw tree.

Phenolic Profiles, Antioxidant and Antimicrobial Activities of Pawpaw Pulp (Asimina triloba [L.] Dunal) at Different Ripening Stages.

In this study, the phenolic components, as well as the antioxidant and antimicrobial activities, of the ripe and unripe fruit of pawpaw (Asimina triloba [L.] Dunal) extracted using five different solvents (distilled water, 95% methanol, 80% methanol, 95% ethanol, and 80% ethanol) were analyzed. The total phenolic content and total flavonoid content were the highest in the 95% ethanol (149.50 mg CAE/g) and 80% ethanol (5.62 mg RE/g) extracts of the unripe fruit, respectively. Analysis of 17 phenolic compounds in pawpaw extracts revealed that epigallocatechin, epicatechin, and p-coumaric acid were the as major compounds, and the amounts of all components significantly decreased with the ripening (P < 0.05). In all antioxidant assays, the 95% ethanol extract of the unripe fruit showed the highest antioxidant activity (EC50 0.22 to 0.93 mg/mL). The pawpaw extracts were more sensitive against Corynebacterium xerosis and Clostridium perfringens. In particular, the 95% ethanol extract of the ripe fruit notably inhibited C. xerosis growth, with minimum inhibitory concentration of 1.56 mg/mL. These results showed that the unripe fruit of pawpaw has abundant phenolic compounds and superior antioxidant activity, and that the 95% ethanol extract of the ripe fruit shows strong inhibitory activity against various microorganisms. Therefore, pawpaw fruit can be utilized as an attractive source of nutrients and therapeutic agents. PRACTICAL APPLICATION: In this study, we identified that the unripe fruit of pawpaw is rich in phenolic compounds and shows strong antioxidant activities. The 95% ethanol extract of the ripe fruit showed strong high inhibitory effect against various microorganisms. These results suggest that pawpaw fruit can serve as a source of antioxidants and delay the aging process. In addition, the fruit could also potentially be utilized as a potential antimicrobial agent.

Correlation Between Acetogenin Content and Antiproliferative Activity of Pawpaw (Asimina triloba [L.] Dunal) Fruit Pulp Grown in Korea.

Pawpaw (Asimina triloba [L.] Dunal) is widely cultivated in Korea for its fruit, which contains bioactive compounds, such as acetogenins. In this study, we investigated the acetogenin content and antiproliferative activity of pawpaw fruit pulp against various cancer cell lines and evaluated the relationship between these two variables at different maturation stages. Unripe fruit had higher antiproliferative activity than ripe fruit, and the activity level depended on acetogenin content. In addition, the presence of specific acetogenins was related to inhibition of certain cancer cell types. The unripe fruit methanol and ethanol extracts (URFM and URFE, respectively) that were rich in acetogenins strongly inhibited the growth of HT-1080, HeLa, and AGS cells by >50% at concentrations of less than 115 µg/mL. These findings indicate that URFM and URFE have therapeutic potential for the treatment of cancer, and our study establishes a basis for further mechanistic studies of the antiproliferative activity of pawpaw fruit. However, it is necessary to further study the anticancer activity of acetogenins from pawpaw fruit using in vivo activity approaches. PRACTICAL APPLICATION: Pawpaw (Asimina triloba) contains acetogenins that can inhibit the growth of cancer cells. In our study, we demonstrate that the antiproliferative activity is higher in unripe than in ripe fruit and depends on acetogenin content. Our results indicate that the extract of unripe pawpaw fruit has value not only as a functional food, but has therapeutic potential for the treatment of cancer as a naturally derived substance that may be less toxic than conventional chemotherapy drugs.

Antioxidant Activities and Phenolic Compounds of Several Tissues of Pawpaw (Asimina triloba [L.] Dunal) Grown in Korea.

Pawpaw (Asimina triloba [L.] Dunal) possesses antioxidant compounds and strong inhibitors of cancer cells, and is widely cultivated in North America, Canada, and Korea. We analyzed the total phenolic and total flavonoid contents (TPC and TFC, respectively) of pawpaw plants grown in Korea and the antioxidant activities of their roots, twigs, leaves, and fruit with respect to 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity, 2,2'-azino-bis diammonium salt (ABTS) radical scavenging activity, ferrous (Fe2+ ) chelating ability, and nitrite scavenging activity. Pearson's correlation analyses revealed a linear correlation between TPC and antioxidant activities (r2 >0.69). Root methanol extracts had higher TPC and antioxidant activities than other extracts, which was also consistent with those from the phenolic compounds found in those extracts. Therefore, antioxidant activities seem to depend on the TPC of each pawpaw tissue and pawpaw roots might be useful as a natural source of natural antioxidants.

Changes in the phenolic compounds and antioxidant activities of mustard leaf (Brassica juncea) kimchi extracts during different fermentation periods.

This study was conducted to investigate the changes in the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activities of 80% methanol and water extracts from mustard leaf kimchi during different fermentation periods. The methanol extract exhibited higher TPC and TFC than the water extract. Both extracts from kimchi fermented for two months showed the highest antioxidant effects against the scavenging activities of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radicals and 2,2-azino-bis diammonium salt (ABTS) radicals. Moreover, the methanol extract from kimchi fermented for two months showed the highest nitrite scavenging activity. The highest metal (Fe2+) chelating effect of the methanol extract and water extract was observed after three months and one month, respectively. Caffeic acid showed the highest increase with fermentation. These findings suggest that the antioxidant activities of kimchi depend on the fermentation period. Accordingly, this study provides basic data for improving the antioxidant activity of mustard leaf kimchi through the establishment of their fermentation period.

Fatty acid compositions of Taenia solium metacestode and its surrounding tissues.

Fatty acids (FAs) are the main energy sources of living organisms and are the major components of cellular and organelle membranes. Their compositions also affect the flexibility/rigidity of cells and cell vitality. The Taenia solium metacestode (TsM) causes neurocysticercosis (NC), which is one of the most common helminthic infections of the central nerve system. We investigated the FA composition of the cyst fluid (CF) and parenchyma of the TsM, together with those of the granuloma and swine tissue surrounding the granuloma. The FA fractions of the TsM CF and swine tissue showed a composition and proportional contents comparable to each other, in which C18:0 (stearic acid), C18:1n9c (oleic acid), C20:4 (arachidonic acid) and C16:0 (palmitic acid) constituted the major fractions. However, the relative amount of individual FAs of the TsM parenchyma and granuloma differed from those of TsM CF and swine tissue, which contained enriched C16:0 and a lower amount of C20:4. Saturated FAs were the major constituents in parenchyma and granuloma, 50.4% and 46.1%, respectively. Conversely, monounsaturated FAs were the major constituents of CF and swine tissue, 38.7% and 40.3%, respectively. Our results strongly suggest that host-derived FAs might translocate across the parasite syncytial membrane and be stored in the CF.

Butanol extract of Ecklonia cava prevents production and aggregation of beta-amyloid, and reduces beta-amyloid mediated neuronal death.

Beta-amyloid (Aß) is a major pathogenic peptide for Alzheimer's disease (AD) and is generated by the processing of amyloid precursor protein (APP). The Aß monomers aggregate into oligomeric and fibrillar forms which have been implicated as the toxic species inducing the neuronal dysfunction. Brown algae Ecklonia cava is known for its anti-oxidant and anti-inflammatory functions. Therefore, we tested the effect of E. cava extract on the production and aggregation of Aß peptides. The butanol extract of E. cava reduced Aß secretion from HEK293 cells expressing APP with Swedish mutation and increased soluble APPα and C-terminal fragment-α (CTFα), of which activity was similar to BACE (ß-site of APP cleaving enzyme) inhibitors. Furthermore, the extract inhibited Aß oligomerization, particularly mid-size oligomer formation, confirmed by the ultrastructural morphology. Congo red, thioflavin T assays, and electron microscopy showed that the extract inhibited Aß fibril formation effectively. Finally, the extract protected primary cortical neurons from various Aß-induced cell deaths, especially oligomer-induced death. Although further study is needed to test the effectiveness of the extract in vivo, our results demonstrate, for the first time, that the butanol extract of E. cava could be used as an anti-Aß agent for AD therapeutics.

Characterization of an extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase from Pseudomonas alcaligenes LB19.

An extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase from an isolate, Pseudomonas alcaligenes LB19, was purified to electrophoretic homogeneity by hydrophobic interaction chromatography using Octyl-Sepharose CL-4B and gel permeation chromatography using Sephadex G-150. The molecular mass of the enzyme, which consisted of a single polypeptide chain, was approximately 27.6 kDa. The pI value of the enzyme was estimated to be 5.7, and its maximum activity was observed at pH 9.0 and 45 degreesC. The enzyme was significantly inactivated by EDTA and phenylmethylsulfonyl fluoride (PMSF) but insensitive to dithiothreitol. It was also markedly inhibited by 0.1% Tween 80 and 0.05% Triton X-100. The purified enzyme could hydrolyze various types of bacterial aliphatic and aromatic MCL-PHAs but not poly(3-hydroxybutyrate), polycaprolactone, and poly(L-lactide). Biodegradation rates of the aromatic MCL-PHAs were significantly lower than those of the aliphatic MCL-PHAs, regardless of the compositions and types of aromatic substituents. It was able to hydrolyze medium-chain-length p-nitrophenylalkanoates more efficiently than the shorter-chain forms. The main hydrolysis products of poly(3-hydroxynonanoate) were identified as monomer units. The results demonstrated in this study suggest that the MCL-PHA depolymerase from P. alcaligenes LB19 is a distinct enzyme, which are different from those of other MCL-PHA degrading bacteria in its quaternary structure, pI value, sensitivity to EDTA and PMSF, and hydrolysis products of MCL-PHA.

Characterization of an extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase from Streptomyces sp. KJ-72.

A bacterial strain capable of degrading medium-chain-length polyhydroxyalkanoates (MCL-PHAs) was isolated from a soil sample. This organism, which was identified as Streptomyces sp. KJ-72, secreted MCL-PHA depolymerase into the culture fluid only when it was cultivated on MCL-PHAs. The extracellular MCL-PHA depolymerase of the organism was purified to electrophoretic homogeneity by ion exchange column chromatography and gel filtration. The enzyme consisted of a monomeric subunit having a molecular mass of 27.1 kDa and isoelectric point of 4.7. The maximum activity was observed at pH 8.7 and 50 degrees C. The enzyme was sensitive to N-bromosuccinimide and acetic anhydride, indicating the presence of tryptophan and lysine residues in the catalytic domain. The enzyme was able to hydrolyze various chain-length p-nitrophenyl esters of fatty acids and polycaprolactone as well as various types of MCL-PHAs. However, lipase activity of the enzyme was not detected. The main hydrolysis product of poly(3-hydroxyheptanoate) was identified to be the dimer of 3-hydroxyheptanoate.