A Novel Fixation Method for Variable-Sized Endoscopic Submucosal Dissection Specimens: An In Vitro Animal Experiment

Background

Endoscopic submucosal dissection is considered a curative and minimally invasive treatment for early gastric cancer; however, precise pathologic assessment of resected specimens is required to develop further treatment plans. Human error during specimen handling can affect objective assessment of resected specimens. In this study, we investigated whether a novel tissue fixation device offered more objective and standardized pathologic evaluation than conventional manual tissue fixation.

Methods

We developed a novel tissue fixation device for endoscopic submucosal dissection specimens. Two circular tissue samples 2, 3, and 4 cm in diameter were obtained from the body of 45 porcine stomachs. One specimen sample was placed in a fixation device; the other was manually fixed on corkboard. We used a pressure indicator to ensure constant pressure in the resected specimens in the fixation device. We measured submucosal diameter and thickness after 24 hr.

Results

The diameters for 2, 3, and 4 cm resected tissue samples were 23.85, 32.30, and 45.0 mm and 21.0, 32.0, and 44.50 mm for the fixation device and manual pinning groups, respectively. The submucosal thicknesses in the fixation device group were 397.09, 381.43, and 415.51 μm and 393.76, 529.69, and 603.82 μm by manual pinning for 2, 3, and 4 cm tissue samples, respectively. Analysis of standard deviation revealed that the submucosal thickness in the manual fixation group was much more variable than in the fixation device group (p = 0.012, 0.042, and 0.001 for 2, 3, and 4 cm tissue specimens, respectively; Fligner-Killeen test of homogeneity of variances).

Conclusions

Among variously sized resected tissue specimens, submucosal thicknesses were more variable in the conventional fixation group, while the thicknesses were comparatively consistent in the fixation device group. After endoscopic submucosal dissection, pathologic preparation using this fixation device could offer more objective assessment of specimens.     

Bacteriorhodopsin production by cell recycle culture of Halobacterium halobium

When Halobacterium halobium R1 was cultured with cell recycle in a bioreactor equipped with an external hollow fiber membrane unit, the cell and bacteriorhodopsin concentrations reached in 10 days were 30.3 g cell dry weight/l and 282 mg/l, respectively. The productivity of bacteriorhodopsin (1.15 mg/l·h) was much higher than that (0.16 mg/l·h) obtained by typical batch fermentation. © Rapid Science Ltd. 1998     

Pseudozyma jejuensis sp. nov., a novel cutinolytic ustilaginomycetous yeast species that is able to degrade plastic waste

An ustilaginomycetous anamorphic yeast, isolated from orange leaves on Jeju island in South Korea, represents a novel Pseudozyma species according to morphologic and physiologic findings and molecular taxonomic analysis using the D1/D2 domains of the large subunit (26S) rRNA gene and the internally transcribed spacer (ITS) 1+2 regions. The name Pseudozyma jejuensis sp. nov. is proposed for this novel species, with OL71(T) (=KCTC 17482(T)=CBS 10454(T)) as type strain. In the present study, we have also demonstrated that Pseudozyma jejuensis OL71 is capable of producing cutinase and degrading polycaprolactone. These results suggest that Pseudozyma jejuensis or its cutinase may be useful for the biological degradation of plastic waste.     

Complete genome sequence of Klebsiella oxytoca KCTC 1686, used in production of 2,3-butanediol

Here we report the full genome sequence of Klebsiella oxytoca KCTC 1686, which is used in production of 2,3-butanediol. The KCTC 1686 strain contains 5,974,109 bp with G+C content of 56.05 mol% and contains 5,488 protein-coding genes and 110 structural RNAs.     

Ethanol extract of paeonia suffruticosa Andrews (PSE) induced AGS human gastric cancer cell apoptosis via fas-dependent apoptosis and MDM2-p53 pathways

ABSTRACT: BACKGROUND: The root bark of Paeonia suffruticosa Andrews (PSE), also known as Moutan Cortex, has been widely used in Asia to treat various diseases. The molecular mechanisms by which PSE exerts its anti-oxidant and anti-inflammatory activities are well known, but its anti-cancer activity is not yet well understood. Here, we present evidence demonstrating that PSE can be used as a potent anti-cancer agent to treat gastric cancer. METHODS: The effects of the ethanol extract of PSE on cell proliferation were determined using an MTT (1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan) assay. Cell cytotoxicity induced by the PSE extact is measured using an LDH leakage assay. Flow cytometry was used to analyze the cell cycle and to measure the subG0/G1 apoptotic cell fraction. Apoptosis induced by the PSE extact is also examined using a DNA fragmentation assay. Western blot analysis is used to measure the levels of apoptotic proteins such as Fas receptor, caspase-8, caspase-3, PARP, Bax, Bcl-2, MDM2, and p53. RESULTS: This study demonstrated that treating AGS cells with the PSE extact significantly inhibited cell proliferation and induced cytotoxicity in a dose- and time-dependent manner. The PSE extract also induced apoptosis in AGS cells, as measured by flow cytometry and a DNA fragmentation assay. We found that the PSE extract induced apoptosis via the extrinsic Fas-mediated apoptosis pathway, which was concurrent with the activation of caspases, including caspase-8 and caspase-3, and cleavage of PARP. The MDM2-p53 pathway also played a role in the apoptosis of AGS cells that was induced by the PSE extract. CONCLUSIONS: These results clearly demonstrate that the PSE extact displays growth-suppressive activity and induces apoptosis in AGS cells. Our data suggest that the PSE extact might be a potential anti-cancer agent for gastric cancer.     

Synthesis of 10-substituted triazolyl artemisinins possessing anticancer activity via Huisgen 1,3-dipolar cylcoaddition

Most of the 10-substituted triazolylartemisinin synthesized via the Huisgen 1,3-dipolar cylcoaddition of diastereomeric 10-azidoartemisinin (5, 6, and 7) with various alkynes (a–h) exhibit strong growth inhibition activity, even at sub-micromolar concentrations, against various cancer cell lines such as DLD-1, U-87, Hela, SiHa, A172, and B16. In particular, 10b and 10f showed a highly strong cytotoxicity.     

1,2,3,4,6-penta-O-galloyl-β-d-glucose: A cholinesterase inhibitor from Terminalia chebula

The search for a novel pharmacotherapy from medicinal plants for neurodegenerative disorders has significantly advanced. Therefore, the present study was performed to evaluate the anticholinesterase activities of one hundred medicinal plants in Korea, where Terminalia chebula (T. chebula) fruits showed significant acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitions. Further bioassay monitored phytochemical exploration led to the isolation of 1,2,3,4,6-penta-O-galloyl-β-d-glucose (compound 1), which showed significant AChE and BChE inhibitory effects with IC₅₀ values of 29.9 ± 0.3 µM and 27.6 ± 0.2 µM, respectively. The inhibitory effect of compound 1 towards acetylcholinesterase was also evaluated using TLC and compared with tacrine as the positive control; the positive effect was confirmed. Furthermore, compound 1 also displayed strong antioxidant activity by the FRAP assay (IC₅₀ = 4.6 ± 0.2 µM). In conclusion, compound 1 may prove to be a potential natural anti-Alzheimer source based on noteworthy AChE and BChE inhibitions, and strong antioxidant activity.     

Preparative separation of chlorogenic acid by centrifugal partition chromatography from highbush blueberry leaves (Vaccinium corymbosum L.)

Introduction – Blueberries (genus Vaccinium) have gained worldwide focus because of the high anthocyanin content of their fruits. In contrast, the leaves of blueberry have not attracted any attention, even though they contain large quantities of chlorogenic acid, a strong antioxidant compound. Objective – The aim of this investigation was the quantification and preparative isolation of chlorogenic acid (5‐caffeoylquinic acid, 5‐CQA) from blueberry leaves using a new separation scheme, centrifugal partition chromatography (CPC). Methodology – A water fraction containing a high concentration of 5‐CQA (14.5% of dry weight extract) was obtained by defatting a crude methanol extract from blueberry leaves. CPC was applied to isolate 5‐CQA from this water fraction using a two‐phase solvent system of ethyl acetate–ethanol–water at a volume ratio 4:1:5 (v/v/v). The flow‐rate of mobile phase was 2 mL/min with the ascending mode while rotating at 1200 rpm. The eluate was monitored at 330 nm. The structure of chlorogenic acid in the CPC fraction was confirmed with HPLC, UV, ESI/MS and NMR spectra. Results – The HPLC chromatogram showed that the fractions collected by CPC contained chlorogenic acid with 96% purity based on peak area percentage. The total amount of chlorogenic acid isolated from 0.5 g of a water fraction was 52.9 mg, corresponding to 10.6% of the water fraction. The isolated compound was identified successively as 5‐CQA with MS (parent ion at m/z 355.1 [M + H]⁺) and ¹H NMR spectra [caffeoyl moiety in the down field (δ 6.0–8.0 ppm) and quinic acid moiety in the up field (δ 2.0–5.5 ppm)]. Conclusion – 5‐CQA was successfully isolated from blueberry leaves by the CPC method in a one‐step procedure, indicating a further potential use for blueberry leaves. Copyright © 2010 John Wiley & Sons, Ltd.