Anticancer Activity of Linalool Terpenoid: Apoptosis Induction and Cell Cycle Arrest in Prostate Cancer Cells

Purpose: To evaluate the anticancer activity of linalool against human prostate cancer (DU145) cells.Methods: The anticancer activity of linalool against DU145 cancer cells was evaluated by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry, using propidium iodide and Annexin V-FITC, was applied to study apoptosis and cell cycle phase distribution. Inverted light microscopy was used to study the effect of linalool on cell morphology and apoptotic body formation in DU145 cells while gel electrophoresis was employed to evaluate the effect of linalool on DNA fragmentation.Results: Linalool induced a dose-dependent as well as time-dependent growth inhibitory effect on DU145 prostate cancer cells. It induced sub-G1 phase growth arrest which led to increase in sub-G0/G1 cell population after treatment with increasing doses of linalool. DNA ladder appeared to be more evident with increasing linalool concentration. However, no DNA fragments were observed in the control groups. It was observed that 4.36, 11.54, 21.88 and 15.54 % of the cells underwent early apoptosis after treatment with 0 (no linalool treatment), 20, 40, and 80 μM of linalool, respectively. Compared to control cells, linalool treatment resulted in the appearance of cell shrinkage along with membrane blebbing which are characteristic features of cell apoptosis.Conclusion: The findings of this study indicate that linalool can be developed as a plant-based chemotherapeutic agent against prostate cancerKeywords: Prostate cancer, Linalool, Chemotherapy, Cell cycle, Apoptosis, DNA fragmentation, Sub- G1 phase growt

Global small RNA analysis in fast-growing Arabidopsis thaliana with elevated concentrations of ATP and sugars

BACKGROUND: In higher eukaryotes, small RNAs play a role in regulating gene expression. Overexpression (OE) lines of Arabidopsis thaliana purple acid phosphatase 2 (AtPAP2) were shown to grow faster and exhibit higher ATP and sugar contents. Leaf microarray studies showed that many genes involved in microRNAs (miRNAs) and trans-acting siRNAs (tasiRNAs) biogenesis were significantly changed in the fast-growing lines. In this study, the sRNA profiles of the leaf and the root of 20-day-old plants were sequenced and the impacts of high energy status on sRNA expression were analyzed. RESULTS: 9-13 million reads from each library were mapped to genome. miRNAs, tasiRNAs and natural antisense transcripts-generated small interfering RNAs (natsiRNAs) were identified and compared between libraries. In the leaf of OE lines, 15 known miRNAs increased in abundance and 9 miRNAs decreased in abundance, whereas in the root of OE lines, 2 known miRNAs increased in abundance and 9 miRNAs decreased in abundance. miRNAs with increased abundance in the leaf and root samples of both OE lines (miR158b and miR172a/b) were predicted to target mRNAs coding for Dof zinc finger protein and Apetala 2 (AP2) proteins, respectively. Furthermore, a significant change in the miR173-tasiRNAs-PPR/TPR network was observed in the leaves of both OE lines. CONCLUSION: In this study, the impact of high energy content on the sRNA profiles of Arabidopsis is reported. While the abundance of many stress-induced miRNAs is unaltered, the abundance of some miRNAs related to plant growth and development (miR172 and miR319) is elevated in the fast-growing lines. An induction of miR173-tasiRNAs-PPR/TPR network was also observed in the OE lines. In contrast, only few cis- and trans-natsiRNAs are altered in the fast-growing lines.published_or_final_versio

High-Tc ramp-type Josephson junctions with a continually graded Y1–xPrxBa2Cu3Oy barrier

High-Tc Josephson junctions with a graded barrier have been prepared by using a composite target. Such a barrier is synthesized by utilizing Y1–xPrxBa2Cu3Oy with a continually graded concentration of Pr, in which no lattice mismatch and other incompatible problems take place. The structural interfaces are absent in the weak link region and Josephson coupling occurs at the naturally formed superconducting/normal interfaces within the Y1–xPrxBa2Cu3Oy layer. Thus, it can significantly enhance the reproducibilty and performance of these junctions. The temperature dependences of the barrier thickness and Josephson were also studied. © 2001 American Institute of Physics.published_or_final_versio

Development of stable marker-free nuclear transformation strategy in the green microalga Chlorella vulgaris

Although microalgae have valuable resources with great necessity in many biotechnological applications, no tools have been developed yet for a stable genetic transformation without antibiotic marker genes in these organisms. Chlorella is one of the most useful genus for biotechnological applications. The transfer of foreign DNA (vector or linear DNA cassette) into Chlorella by electroporation has very low stability and it is hard to screen the transformants without antibiotic marker genes. However, the marker genes have some disadvantages to host cells. To avoid the negative effects caused by the marker genes, we tried to develop a stable marker-free nuclear transformation system in Chlorella. For this, linear gene expression cassettes (LGEC) were constructed with functional domains, which are responsible for transformation, of SV40 large T antigen. The LGECs were transferred into Chlorella via electroporation and durability of the LGECs were confirmed in transgenic Chlorella. Transcription levels of the LGECs were also determined at different cell cycle sates. The LGECs integrated into the chromosomal DNA of Chlorella were stably replicated and were expressed successfully at G0-, G1-, and G2-phases. This study presents a stable marker-free nuclear transformation system with potential for biotechnological applications.Key words: Chlorella vulgaris, marker-free nuclear transformation, SV40 large T antigen, microalga

Studies on Anti-Hepatoma Effect of Gan-Ai-Xiao Decoction

Purpose: To explore the anti-hepatoma effect of Gan-Ai-Xiao Decoction (GAXD), a folk remedy.Methods: High performance liquid chromatography (HPLC) was used to identify the major chemical components of GAXD ethanol extract (EE). The cytotoxic effect of GAXD EE against HepG2 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Flow cytometry and Western blot were used to study the effect of GAXD EE on apoptosis and apoptotic proteins (Bcl-2, Bax and caspase-3) in HepG2 cells. Xenograft assay was used to evaluate the anti-hepatoma effect of GAXD EE in vivo.Results: Four components were identified in GAXD EE by HPLC. The results of MTT and flow cytometry assays indicated that GAXD EE significantly reduced HepG2 cells viability (p < 0.05) and induced its apoptosis. The results of Western blot assay suggested that GAXD EE down-regulated the expression of anti-apoptotic protein (Bcl-2) and up-regulated the expression of pro-apoptotic proteins (Bax and caspase-3) in HepG2 cells. Furthermore, the results of xenograft assay showed that GAXD EE significantly inhibited the growth of HepG2 cells-induced tumor (p < 0.05) without any effect on the body weight of nude mice.Conclusion: GAXD has anti-hepatoma activity, and the mechanism is associated with apoptosis.Keywords: Gan-Ai-Xiao Decoction, Anti-hepatoma, Flow Cytometry, Western Blot, Xenograft, HepG2 cells, Apoptosi

A comparison of acidic and enzymatic hydrolysis of rutin

Rutin and its hydrolysis products (isoquercitrin and quercetin) are widely used as important materials in food and pharmaceutical industry. In this study, the effects of various acids and enzymes as catalysts on the hydrolysis reaction of rutin were studied. In comparison with acidic and enzymatic catalysis of rutin, the research results indicated that there was a sharp difference in the selectivity of hydrolysis product between the methods. When 2.5% H3PO4, 1% HCl and 0.5% H2SO4 were used as catalysts, transformation yields of isoquercitin hydrolyzed from rutin were 9.60, 0.69 and 1.25%, but those of quercetin were 11.13, 100 and 2.57%, respectively. When hesperidinase, snailase and cellulase-T2440 were used as catalysts, transformation yields of isoquercitin hydrolyzed from rutin were 43.21, 3.07 and 0.00%, but those of quercetin were 58.10, 96.39 and 30.89%, respectively. In conclusion, the aglycon of rutin was deglycosolated easily under mild acidic hydrolysis conditions at appropriate temperatures, but its secondary glucoside was difficult to be obtained. Contrarily, the prepared isoquercitrin by enzymatic hydrolysis of rutin was preferable to the acidic hydrolysis, especially for hesperidinase.Key words: Rutin, isoquercitrin, quercetin, hydrolysis, acid, enzym

Simple models of the chemical field around swimming plankton

Background. Cervical cancer is the fourth most common cancer in women, and we recently reported human leukocyte antigen (HLA) alleles showing strong associations with cervical neoplasia risk and protection. HLA ligands are recognized by killer immunoglobulin-like receptors (KIRs) expressed on a range of immune cell subsets, governing their proinflammatory activity. We hypothesized that the inheritance of particular HLA-KIR combinations would increase cervical neoplasia risk. Methods. Here, we used HLA and KIR dosages imputed from single-nucleotide polymorphism genotype data from 2143 cervical neoplasia cases and 13 858 healthy controls of European decent. Results. The following 4 novel HLA alleles were identified in association with cervical neoplasia, owing to their linkage disequilibrium with known cervical neoplasia-associated HLA-DRB1 alleles: HLA-DRB3*9901 (odds ratio [OR], 1.24; P = 2.49 × 10−9), HLA-DRB5*0101 (OR, 1.29; P = 2.26 × 10−8), HLA-DRB5*9901 (OR, 0.77; P = 1.90 × 10−9), and HLA-DRB3*0301 (OR, 0.63; P = 4.06 × 10−5). We also found that homozygosity of HLA-C1 group alleles is a protective factor for human papillomavirus type 16 (HPV16)-related cervical neoplasia (C1/C1; OR, 0.79; P = .005). This protective association was restricted to carriers of either KIR2DL2 (OR, 0.67; P = .00045) or KIR2DS2 (OR, 0.69; P = .0006). Conclusions. Our findings suggest that HLA-C1 group alleles play a role in protecting against HPV16-related cervical neoplasia, mainly through a KIR-mediated mechanism

Anticoagulation Control in Warfarin-Treated Patients Undergoing Cardioversion of Atrial Fibrillation (from the Edoxaban Versus Enoxaparin-Warfarin in Patients Undergoing Cardioversion of Atrial Fibrillation Trial).

In the Edoxaban Versus Enoxaparin-Warfarin in Patients Undergoing Cardioversion of Atrial Fibrillation (ENSURE-AF) study (NCT 02072434), edoxaban was compared with enoxaparin-warfarin in 2,199 patients undergoing electrical cardioversion of nonvalvular atrial fibrillation (AF). In this multicenter prospective randomized open blinded end-point trial, we analyzed patients randomized to enoxaparin-warfarin. We determined time to achieve therapeutic range (TtTR); time in therapeutic range (TiTR); their clinical determinants; relation to sex, age, medical history, treatment, tobacco use, race risk (SAMe-TT2R2) score; and impact on primary end points (composite of stroke, systemic embolic event[SEE], myocardial infarction [MI], and cardiovascular death [CVD] and composite of major + clinically relevant nonmajor bleeding). Among 1,104 patients randomized to enoxaparin-warfarin, 27% were naïve to oral anticoagulants. Mean age was 64.2 ± 11 years and mean congestive heart failure, hypertension, age ≥75 (doubled), diabetes mellitus, prior stroke or transient ischemic attack (doubled), vascular disease, age 65-74, female (CHA2DS2-VASc) score was 2.6. Mean TtTR was 7.7 days (median 7 days) and mean TiTR after reaching an international normalized ratio of 2.0 to 3.0 was 71%. In 695 patients who had an INR 2. On multivariate regression, an independent predictor of extended TtTR was creatinine clearance (p = 0.02). TtTR was marginally related to stroke/SEE/MI/CVD (p = 0.06; odds ratio  0.23, 95% confidence interval 0.02 to 1.17) but not to any bleeding. Independent predictors of TiTR were previous vitamin K antagonist experience (p65, concomitant drugs or alcohol (HAS-BLED) score (p = 0.02). TiTR was related to any bleeding (p = 0.02; odds ratio  0.39, 95% confidence interval 0.16 to 0.88), but not stroke/SEE/MI/CVD. In this cohort of warfarin users with a high TiTR no difference was seen between TtTR and TiTR in relation to SAMe-TT2R2 score. In conclusion, even in this short-term study, TiTR was significantly related to bleeding events

Comparison of Small RNA Profiles of Glycine max and Glycine soja at Early Developmental Stages

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