The cytotoxic effect of usnic acid in malignant melanoma cells with different genomic profiles in the BRAF aspect

Objective: Malignant melanoma (MM) is the most aggressive skin cancer and treatment options are still limited in the late stages, generally accompanied by BRAF mutations. Usnic acid (UA), a well-known traditional lichen metabolite, has a promising and selective antitumoral activity. However, the effects of UA on MM cells with different genomic profiles in the BRAF aspect have not been investigated yet. In this study, we evaluated the effect of UA on BRAFV600E mutated-A2058 and wild-type MeWo cells. Materials and Methods: In the UA-treated cells, viability and cell death analysis were performed by using WST-1 and Annexin-V assays. Then, the death-related morphological changes were visualized by acridine orange (AO)/ethidium bromide (EB) staining. The cell cycle regulatory effect of UA was determined. Finally, time-dependent detection of acidic vesicular organelles (AVOs) was performed by live-cell imaging. Results: While MeWo viability significantly reduced to 53.8% and 28.6%, A2058 viability was detected as 61.3% and 50.3% at 50 and 100 µM UA for 48 h. Thus, MeWo cells were found to be more sensitive to UA. Annexin-V and morphological analysis results showed that UA triggered mainly a vacuole-dependent cell death by the formation of AVOs, instead of apoptosis, in the MM cells. This effect was prominent in A2058 compared to MeWo. UA also slightly triggered apoptosis in MeWo cells. Thus, the cell cycle regulatory effect of UA on MM cells changed based on the cell death type triggered. Conclusions: Our results suggest that UA exerts the cytotoxic effects on MM cells by inducing vacuole-dependent cell death, most probably autophagy, and the UA response of MM cells with a different genomic profile in the BRAF aspect varies

Synthesis of superlow friction carbon films from highly hydrogenated methane plasmas.

In this study, we investigated the friction and wear performance of diamondlike carbon films (DLC) derived from increasingly hydrogenated methane plasmas. The films were deposited on steel substrates by a plasma-enhanced chemical vapor deposition process at room temperature and the tribological tests were performed in dry nitrogen. Tests results revealed a close correlation between the hydrogen in source gas plasma and the friction and wear coefficients of the DLC films. Specifically, films grown in plasmas with higher hydrogen-to-carbon ratios had much lower friction coefficients and wear rates than did films derived from source gases with lower hydrogen-to-carbon ratios. The lowest friction coefficient (0.003) was achieved with a film derived from 25% methane--75% hydrogen, while a coefficient of 0.015 was found for films derived from pure methane. Similar correlations were observed for wear rates. Films derived from hydrogen-rich plasmas had the least wear, while films derived from pure methane suffered the highest wear. We used a combination of surface analytical methods to characterize the structure and chemistry of the DLC films and worn surfaces

Tribological Behavior of Near-Frictionless Carbon Coatings in High- and Low- Sulfur Diesel Fuels

The sulfur content in diesel fuel has a significant effect on diesel engine emissions, which are currently subject to environmental regulations. It has been observed that engine particulate and gaseous emissions are directly proportional to fuel sulfur content. With the introduction of low-sulfur fuels, significant reductions in emissions are expected. The process of sulfur reduction in petroleum-based diesel fuels also reduces the lubricity of the fuel, resulting in premature failure of fuel injectors. Thus, another means of preventing injector failures is needed for engines operating with low-sulfur diesel fuels. In this study, the authors evaluated a near-frictionless carbon (NFC) coating (developed at Argonne National Laboratory) as a possible solution to the problems associated with fuel injector failures in low-lubricity fuels. Tribological tests were conducted with NFC-coated and uncoated H13 and 52100 steels lubricated with high- and low- sulfur diesel fuels in a high-frequency reciprocating test machine. The test results showed that the NFC coatings reduced wear rates by a factor of 10 over those of uncoated steel surfaces. In low-sulfur diesel fuel, the reduction in wear rate was even greater (i.e., by a factor of 12 compared to that of uncoated test pairs), indicating that the NFC coating holds promise as a potential solution to wear problems associated with the use of low-lubricity diesel fuels

Long term survival after coronary endarterectomy in patients undergoing combined coronary and valvular surgery – a fifteen year experience

<p>Abstract</p> <p>Background</p> <p>Coronary Endarterectomy (CE) in patients undergoing coronary artery bypass graft (CABG) surgery has been shown to be beneficial in those with diffuse coronary artery disease. There are no published data on its role and benefit in patients undergoing more complex operations. We present our experience with CE in patients undergoing valve surgery with concomitant CABG.</p> <p>Materials and methods</p> <p>Between 1989 and 2003, 237 patients underwent CABG with valve surgery under a single surgeon at our institution. Of these, 41 patients needed CE. Data was retrospectively obtained from hospital records and database. Further follow-up was obtained by telephone interview. All variables were analyzed by univariate analysis for significant factors relating to hospital mortality. Morbidity and long term survival was also studied. There were 29 males and 12 females with a mean age of 67.4 ± 8.1 and body mass index of 26.3 ± 3.3. Their mean euroscore was 7.6 ± 3.2 and the log euro score was 12.2 ± 16.1.</p> <p>Results</p> <p>Thirty-two patients were discharged from the intensive therapy unit within 48 hours after surgery. Average hospital stay was 12.7 ± 10.43 days. Thirty day mortality was 9.8%. Six late deaths occurred during the 14 year follow up. Ten year survival was 57.2% (95% CL 37.8%–86.6%). Three of the survivors had Class II symptoms, with one requiring nitrates. None required further percutaneous or surgical intervention. We compared the result with the available mortality figure from the SCTS database.</p> <p>Conclusion</p> <p>Compared to the SCTS database for these patients, we have observed that CE does not increase the mortality in combined procedures. By accomplishing revascularization in areas deemed ungraftable, we have shown an added survival benefit in this group of patients.</p

Early results of coronary artery bypass grafting with coronary endarterectomy for severe coronary artery disease

<p>Abstract</p> <p>Background</p> <p>Despite the existence of controversial debates on the efficiency of coronary endarterectomy (CE), it is still used as an adjunct to coronary artery bypass grafting (CABG). This is particularly true in patients with endstage coronary artery disease. Given the improvements in cardiac surgery and postoperative care, as well as the rising number of elderly patient with numerous co-morbidities, re-evaluating the pros and cons of this technique is needed.</p> <p>Methods</p> <p>Patient demographic information, operative details and outcome data of 104 patients with diffuse calcified coronary artery disease were retrospectively analyzed with respect to functional capacity (NYHA), angina pectoris (CCS) and mortality. Actuarial survival was reported using a Kaplan-Meyer analysis.</p> <p>Results</p> <p>Between August 2001 and March 2005, 104 patients underwent coronary artery bypass grafting (CABG) with adjunctive coronary endarterectomy (CE) in the Department of Thoracic-, Cardiac- and Vascular Surgery, University of Goettingen. Four patients were lost during follow-up. Data were gained from 88 male and 12 female patients; mean age was 65.5 ± 9 years. A total of 396 vessels were bypassed (4 ± 0.9 vessels per patient). In 98% left internal thoracic artery (LITA) was used as arterial bypass graft and a total of 114 vessels were endarterectomized. CE was performed on right coronary artery (RCA) (n = 55), on left anterior descending artery (LAD) (n = 52) and circumflex artery (RCX) (n = 7). Ninety-five patients suffered from 3-vessel-disease, 3 from 2-vessel- and 2 from 1-vessel-disease. Closed technique was used in 18%, open technique in 79% and in 3% a combination of both. The most frequent endarterectomized localization was right coronary artery (RCA = 55%). Despite the severity of endstage atherosclerosis, hospital mortality was only 5% (n = 5). During follow-up (24.5 ± 13.4 months), which is 96% complete (4 patients were lost caused by unknown address) 8 patients died (cardiac failure: 3; stroke: 1; cancer: 1; unknown reasons: 3). NYHA-classification significantly improved after CABG with CE from 2.2 ± 0.9 preoperative to 1.7 ± 0.9 postoperative. CCS also changed from 2.4 ± 1.0 to 1.5 ± 0.8</p> <p>Conclusion</p> <p>Early results of coronary endarterectomy are acceptable with respect to mortality, NYHA & CCS. This technique offers a valuable surgical option for patients with endstage coronary artery disease in whom complete revascularization otherwise can not be obtained. Careful patient selection will be necessary to assure the long-term benefit of this procedure.</p

The ϕ6 Cystovirus Protein P7 Becomes Accessible to Antibodies in the Transcribing Nucleocapsid: A Probe for Viral Structural Elements

Protein P7 is a component of the cystovirus viral polymerase complex. In the unpackaged procapsid, the protein is situated in close proximity to the viral directed RNA polymerase, P2. Cryo-electron microscopy difference maps from the species ϕ6 procapsid have demonstrated that P7 and P2 likely interact prior to viral RNA packaging. The location of P7 in the post-packaged nucleocapsid (NC) remains unknown. P7 may translocate closer to the five-fold axis of a filled procapsid but this has not been directly visualized. We propose that monoclonal antibodies (Mabs) can be selected that serve as probe- reagents for viral assembly and structure. A set of Mabs have been isolated that recognize and bind to the ϕ6 P7. The antibody set contains five unique Mabs, four of which recognize a linear epitope and one which recognizes a conformational epitope. The four unique Mabs that recognize a linear epitope display restricted utilization of Vκ and VH genes. The restricted genetic range among 4 of the 5 antibodies implies that the antibody repertoire is limited. The limitation could be the consequence of a paucity of exposed antigenic sites on the ϕ6 P7 surface. It is further demonstrated that within ϕ6 nucleocapsids that are primed for early-phase transcription, P7 is partially accessible to the Mabs, indicating that the nucleocapsid shell (protein P8) has undergone partial disassembly exposing the protein’s antigenic sites

Structural Studies of the Tandem Tudor Domains of Fragile X Mental Retardation Related Proteins FXR1 and FXR2

Expansion of the CGG trinucleotide repeat in the 5′-untranslated region of the FMR1, fragile X mental retardation 1, gene results in suppression of protein expression for this gene and is the underlying cause of Fragile X syndrome. In unaffected individuals, the FMRP protein, together with two additional paralogues (Fragile X Mental Retardation Syndrome-related Protein 1 and 2), associates with mRNA to form a ribonucleoprotein complex in the nucleus that is transported to dendrites and spines of neuronal cells. It is thought that the fragile X family of proteins contributes to the regulation of protein synthesis at sites where mRNAs are locally translated in response to stimuli.Here, we report the X-ray crystal structures of the non-canonical nuclear localization signals of the FXR1 and FXR2 autosomal paralogues of FMRP, which were determined at 2.50 and 1.92 Å, respectively. The nuclear localization signals of the FXR1 and FXR2 comprise tandem Tudor domain architectures, closely resembling that of UHRF1, which is proposed to bind methylated histone H3K9.The FMRP, FXR1 and FXR2 proteins comprise a small family of highly conserved proteins that appear to be important in translational regulation, particularly in neuronal cells. The crystal structures of the N-terminal tandem Tudor domains of FXR1 and FXR2 revealed a conserved architecture with that of FMRP. Biochemical analysis of the tandem Tudor doamins reveals their ability to preferentially recognize trimethylated peptides in a sequence-specific manner

Collagen fleeces do not improve colonic anastomotic strength but increase bowel obstructions in an experimental rat model

To investigate whether a collagen fleece kept in place by fibrin glue might seal off a colorectal anastomosis, provide reinforcement, and subsequently improve anastomotic healing. Wistar rats underwent a 1-cm left-sided colonic resection followed by a 4-suture end-to-end anastomosis. They were then randomly assigned to one of three treatment groups: no additional intervention (control, n = 20), the anastomosis covered with fibrin glue (fibrin glue, n = 20), the anastomosis covered with a collagen fleece, kept in place with fibrin glue (collagen fleece, n = 21). At either 3 or 7 days follow-up, anastomotic bursting pressure was measured and tissue was obtained for histology and collagen content assessment after which animals were sacrificed. Three rats in the control (15%), three in the fibrin glue (15%), and one in the collagen group (4.8%) died due to anastomotic complications (P = 0.497). Anastomotic bursting pressures were not significantly different between groups at 3 and 7 days follow-up (P = 0.659 and P = 0.427, respectively). However, bowel obstructions occurred significantly more often in the collagen group compared to the control group (14/21 vs. 3/20, P = 0.003). Collagen contents were not different between groups, but histology showed a more severe inflammation in the collagen group compared to the other groups at both 3 and 7 days follow-up. A collagen fleece kept in place by fibrin glue does not improve healing of colonic anastomoses in rats. Moreover, this technique induces significantly more bowel obstructions in rats, warranting further study before being translated to a clinical settin

Using leap motion to investigate the emergence of structure in speech and language

© 2016, The Author(s). In evolutionary linguistics, experiments using artificial signal spaces are being used to investigate the emergenceof speech structure. These signal spaces need to be continuous, non-discretized spaces from which discrete unitsand patterns can emerge. They need to be dissimilar from—but comparable with—the vocal tract, in order tominimize interference from pre-existing linguistic knowledge, while informing us about language. This is a hardbalance to strike. This article outlines a new approach that uses the Leap Motion, an infrared controller that canconvert manual movement in 3d space into sound. The signal space using this approach is more flexible than signalspaces in previous attempts. Further, output data using this approach is simpler to arrange and analyze. Theexperimental interface was built using free, and mostly open- source libraries in Python. We provide our sourcecode for other researchers as open source