Libertellenone H,a Natural Pimarane Diterpenoid,Inhibits Thioredoxin System and Induces ROS-Mediated Apoptosis in Human Pancreatic Cancer Cells

Libertellenone H (LH), a marine-derived pimarane diterpenoid isolated from arctic fungus Eutypella sp. D-1, has shown effective cytotoxicity on a range of cancer cells. The present study is to explore the anticancer effect of LH on human pancreatic cancer cells and to investigate the intracellular molecular target and underlying mechanism. As shown, LH exhibited anticancer activity in human pancreatic cancer cells by promoting cell apoptosis. Mechanistic studies suggested that LH-induced reactive oxygen species (ROS) accumulation was responsible for apoptosis as antioxidant N-acetylcysteine (NAC) and antioxidant enzyme superoxide dismutase (SOD) antagonized the inhibitory effect of LH. Zymologic testing demonstrated that LH inhibited Trx system but had little effect on the glutathione reductase and glutaredoxin. Mass spectrometry (MS) analysis revealed that the mechanism of action was based on the direct conjugation of LH to the Cys³²/Cys³⁵ residue of Trx1 and Sec⁴⁹⁸ of TrxR, leading to a decrease in the cellular level of glutathione (GSH) and activation of downstream ASK1/JNK signaling pathway. Taken together, our findings revealed LH was a marine derived inhibitor of Trx system and an anticancer candidate.     

Development and evaluation of novel hydrogel for preventing postoperative pancreatic fistula

Pancreatic fistula is a complication that frequently occurs after pancreatic surgery. Although various attempts have been made to prevent pancreatic fistula after pancreatic surgery, no effective methods have been developed thus far. In this study, to prevent the pancreatic fistula, we attempted to prepare the hydrogels consisting of two types of polyvinyl alcohol (PVAs), Poval® and Exceval® by the freezing–thawing (F/T) method. As the concentration and the number of F/T cycles increased, it was revealed that the swelling degree tended to decrease, and tensile strength tended to increase. Especially, Exceval® formed more robust hydrogels than that of Poval® hydrogel prepared by the same conditions. Furthermore, the release behavior of tartrazine from 10% Exceval® hydrogels differed depending on the number of F/T cycles. It was also revealed that the diffusion pattern of the drug in the hydrogel differed along with the release, and the entire amount of tartrazine in the hydrogel was released by 120 min. As nafamostat mesylate was loaded, in vivo study, Pancreatic enzyme values were obtained after 48 h, which allowed comparison of the preventive effects against pancreatic fistula between the untreated group, the Exceval® hydrogel group, and nafamostat mesylate-containing Exceval®-applied group. Furthermore, there was a significant difference between the untreated group and the hydrogel-applied group. Besides, PVA hydrogels prepared by the F/T method could probably absorb pancreatic enzymes. This study found that Exceval® hydrogel prepared by the F/T method was considered a novel hydrogel formulation to prevent pancreatic fistula.     

Immobilized Au nanoparticles on chitosan-biguanidine modified Fe3O4 nanoparticles and investigation of its anti-human lung cancer activity

In current nanoscience bioengineered magnetic nanoparticles (NPs) have come into prominence with considerable impact. These advanced functional materials find outstanding applications in chemical science in catalysis, environmental issues, sensing etc, as well as in biology as drug delivery agent, chemical therapeutics and others. We have been prompted to architect and synthesize a novel Au NP adorned over chitosan-biguanidine polyplex modified core–shell type magnetic nanocomposite (Fe₃O₄/CS-biguanidine/Au NPs). The bioshells facilitate to protect the core ferrite NPs as well as provides stability to the synthesized Au NPs by capping. The material was characterized using different analytical techniques like Fourier Transformed Infra-Red spectroscopy (FT-IR), Inductively Coupled Plasma-Optical Emission Microscopy (ICP-OES), Field Emission Scanning Electron Microscopy (FE-SEM), Energy Dispersive X-ray spectroscopy (EDX), Transmission Electron Microscopy (TEM), Vibrating Sample Magnetometer (VSM) and X-ray Diffraction (XRD) studies. We explored the biological application of the nanocomposite in determining cytotoxicity of three adenocarcinoma cell lines (PC-14, LC-2/ad, HLC-1) through the MTT assay. The material showed very good activity by exhibiting very low % cell viability over the cell lines dose-dependently. The IC50 of Fe₃O₄/CS-biguanidine/Au NPs were observed 503, 398 and 475 µg/mL respectively against the three cell lines. The best output was observed at a concentration of 1000 µg/mL of catalyst in terms of cytotoxicity and inhibition of lung cancer growth. The anti-cancer potential was found in close relation to their antioxidant potential.     

Collective motions of rigid fragments in protein structures from smoothed electron density distributions

In this work, the Gaussian Network Model (GNM) and Anisotropic Network Model (ANM) approaches are applied to describe the dynamics of protein structure graphs built from calculated promolecular electron density (ED) distribution functions. A first set of analyses is carried out on results obtained from ED maxima calculated at various smoothing levels. A second set is achieved for ED networks whose edges are weighted by ED overlap integral values. Results are compared with those obtained through the classical GNM and ANM approaches applied to networks of C(alpha) atoms. It is shown how the network model and the consideration of crystal packing as well as of the side chains may lead to various improvements dependent upon the structure under study. The selected protein structures are Crambin and Pancreatic Trypsin Inhibitor because of their small size and numerous dynamical data obtained by other authors.     

Stromal Vascular Fraction Loaded Silk Fibroin Mats Effectively Support the Survival of Diabetic Mice after Pancreatic Islet Transplantation

The aim of this study is to assess whether stromal vascular fraction (SVF)‐soaked silk fibroin nonwoven mats (silk‐SVF) can preserve the functionality of encapsulated pancreatic endocrine cells (alginate‐PECs) after transplantation in the subcutaneous tissue of diabetic mice. Silk scaffolds are selected to create an effective 3D microenvironment for SVF delivery in the subcutaneous tissue before diabetes induction: silk‐SVF is subcutaneously implanted in the dorsal area of five healthy animals; after 15 d, mice are treated with streptozotocin to induce diabetes and then alginate‐PECs are implanted on the silk‐SVF. All animals appear in good health, increasing weight during time, and among them, one presents euglycemia until the end of experiments. On the contrary, when PECs are simultaneously implanted with SVF after diabetes induction, mice are euthanized due to suffering. This work clearly demonstrates that silk‐SVF creates a functional niche in subcutaneous tissue and preserves endocrine cell survival and engraftment.     

Effect of Resveratrol Treatment on Human Pancreatic Cancer Cells through Alterations of Bcl-2 Family Members

Pancreatic cancers are among of the most lethal types of neoplasms, and are mostly detected at an advanced stage. Conventional treatment methods such as chemotherapy or radiotherapy often do not bring the desired therapeutic effects. For this reason, natural compounds are increasingly being used as adjuvants in cancer therapy. Polyphenolic compounds, including resveratrol, are of particular interest. The aim of this study is to analyze the antiproliferative and pro-apoptotic mechanisms of resveratrol on human pancreatic cells. The study was carried out on three human pancreatic cancer cell lines: EPP85-181P, EPP85-181RNOV (mitoxantrone-resistant cells) and AsPC-1, as well as the normal pancreatic cell line H6c7. The cytotoxicity of resveratrol in the tested cell lines was assessed by the colorimetric method (MTT) and the flow cytometry method. Three selected concentrations of the compound (25, 50 and 100 µM) were tested in the experiments during a 48-h incubation. TUNEL and Comet assays, flow cytometry, immunocytochemistry, confocal microscopy, real-time PCR and Western Blot analyses were used to evaluate the pleiotropic effect of resveratrol. The results indicate that resveratrol is likely to be anticarcinogenic by inhibiting human pancreatic cancer cell proliferation. In addition, it affects the levels of Bcl-2 pro- and anti-apoptotic proteins. However, it should be emphasized that the activity of resveratrol was specific for each of the tested cell lines, and the most statistically significant changes were observed in the mitoxantrone-resistant cells.     

电化学发光-聚合酶链式反应方法检测胰腺癌中K-ras基因点突变

发展了一种可用于快速检测胰腺癌中K-ras癌基因点突变的电化学发光-聚合酶链式反应(ECL-PCR)分析方法。该法采用三联吡啶钌标记的上游引物和生物素标记的下游引物对目的片段进行PCR扩增;再采用限制性内切酶MvaI对扩增产物进行酶切。由于野生型样品和突变型样品间存在酶切位点的变化,其中只有野生型样品能被切断;通过生物素与链霉亲和素包被的磁珠连接,将生物素标记的DNA片段收集到检测池中,进行电化学发光检测。采用该法对13例胰腺癌组织中的K-ras癌基因第12位密码子进行点突变分析,只需要10μL样品、20min孵育时间和30s采集时间,就可得出其中有12例存在点突变,点突变率为92.3%。本方法操作简便、安全、快速、灵敏,可用于检测任何一种导致限制性内切酶位点改变的基因点突变。     

Sesquiterpene Lactones as Promising Candidates for Cancer Therapy: Focus on Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease which confers to patients a poor prognosis at short term. PDAC is the fourth leading cause of death among cancers in the Western world. The rate of new cases of pancreatic cancer (incidence) is 10 per 100,000 but present a 5-year survival of less than 10%, highlighting the poor prognosis of this pathology. Furthermore, 90% of advanced PDAC tumor present KRAS mutations impacting in several oncogenic signaling pathways, many of them associated with cell proliferation and tumor progression. Different combinations of chemotherapeutic agents have been tested over the years without an improvement of significance in its treatment. PDAC remains as one the more challenging biomedical topics thus far. The lack of a proper early diagnosis, the notable mortality statistics and the poor outcome with the available therapies urge the entire scientific community to find novel approaches against PDAC with real improvements in patients’ survival and life quality. Natural compounds have played an important role in the process of discovery and development of new drugs. Among them, terpenoids, such as sesquiterpene lactones, stand out due to their biological activities and pharmacological potential as antitumor agents. In this review, we will describe the sesquiterpene lactones with in vitro and in vivo activity against pancreatic tumor cells. We will also discuss the mechanism of action of the compounds as well as the signaling pathways associated with their activity.