Automatic Vessel Detection Technique in Laparoscopic Surgical Imaging Including Surgical Instruments.

In this paper, we propose a new technique to recognize vessels in robot-assisted laparoscopic surgery images by using surgical instruments. The proposed method does not require additional hardware or parameter adjustment because it detects blood vessels by using only the color information of the image. The concept of a hessian matrix is used in the HSV color space of the image to detect the edges of the blood vessels. In addition, the histogram equalization technique, clustering technique, and region growing are used to remove the surgical tools. Images of actual robot-assisted laparoscopic surgery videos were used. The processing speed was approximately 0.3 s per frame at 640p and approximately 0.8 s per frame at 1280p. The average recall was 92.67%.

Host Cell Nuclear Localization of Shigella flexneri Effector OspF Is Facilitated by SUMOylation.

When Shigella infect host cells, various effecter molecules are delivered into the cytoplasm of the host cell through the type III secretion system (TTSS) to facilitate their invasion process and control the host immune responses. Among these effectors, the S. flexneri effector OspF dephosphorylates mitogen-activated protein kinases and translocates itself to the nucleus, thus preventing histone H3 modification to regulate expression of proinflammatory cytokines. Despite the critical role of OspF, the mechanism by which it localizes in the nucleus has remained to be elucidated. In the present study, we identified a potential small ubiquitin-related modifier (SUMO) modification site within OspF and we demonstrated that Shigella TTSS effector OspF is conjugated with SUMO in the host cell and this modification mediates the nuclear translocation of OspF. Our results show a bacterial virulence factor can exploit host post-translational machinery to execute its intracellular trafficking.

Activated Carbon Textile via Chemistry of Metal Extraction for Supercapacitors.

Carbothermic reduction in the chemistry of metal extraction (MO(s) + C(s) → M(s) + CO(g)) using carbon as a sacrificial agent has been used to smelt metals from diverse oxide ores since ancient times. Here, we paid attention to another aspect of the carbothermic reduction to prepare an activated carbon textile for high-rate-performance supercapacitors. On the basis of thermodynamic reducibility of metal oxides reported by Ellingham, we employed not carbon, but metal oxide as a sacrificial agent in order to prepare an activated carbon textile. We conformally coated ZnO on a bare cotton textile using atomic layer deposition, followed by pyrolysis at high temperature (C(s) + ZnO(s) → C'(s) + Zn(g) + CO(g)). We figured out that it leads to concurrent carbonization and activation in a chemical as well as mechanical way. Particularly, the combined effects of mechanical buckling and fracture that occurred between ZnO and cotton turned out to play an important role in carbonizing and activating the cotton textile, thereby significantly increasing surface area (nearly 10 times) compared with the cotton textile prepared without ZnO. The carbon textiles prepared by carbothermic reduction showed impressive combination properties of high power and energy densities (over 20-fold increase) together with high cyclic stability.

Autotrophic denitrification performance and bacterial community at biocathodes of bioelectrochemical systems with either abiotic or biotic anodes.

Two-chamber bioelectrochemical systems (BESs) have recently been developed for nitrate removal from nitrate-contaminated water. In this study, we compared the nitrate removal performance of biocathodes of BESs when using abiotic and biotic anodes. Acetate was used as electron donor in BESs with biotic anode, whereas a direct current power supply was used as energy source in BESs with abiotic anode. The nitrogen removal efficiency increased from 18.1% to 43.0% when the voltage supplied to the BES with abiotic anode increased from 0.7 V to 0.9 V, whereas no higher removal efficiency was obtained at a higher supplied voltage (1.1 V). The highest efficiency (78.0%) of autotrophic nitrogen removal was achieved when electron transfer from the biotic anode chamber of BESs was used. Unexpectedly, control of the cathode potential did not enhance nitrate removal in BESs with biotic anode. Special attention was paid to elucidate the differences of bacterial communities catalysing autotrophic denitrification in the biocathodes of BESs with abiotic and biotic anodes. Data from denaturing gradient gel electrophoresis and phylogenetic analysis suggested that denitrification in BESs with abiotic anode could be attributed to Nitratireductor sp., Shinella sp., and Dyella sp., whereas the dominant bacterial denitrifiers in BESs with biotic anode were found to be Pseudomonas sp., Curtobacterium sp., and Aeromonas sp. These results implied that biocathodes of BESs with biotic anode are more efficient than those of BESs with abiotic anode for nitrate removal from nitrate-contaminated water in practical applications.

Hydroquinone diphosphate as a phosphatase substrate in enzymatic amplification combined with electrochemical-chemical-chemical redox cycling for the detection of E. coli O157:H7.

Signal amplification by enzyme labels in enzyme-linked immunosorbent assays (ELISAs) is not sufficient for detecting a low number of bacterial pathogens. It is useful to employ approaches that involve multiple signal amplification such as enzymatic amplification plus redox cycling. An advantageous combination of an enzyme product [for fast electrochemical-chemical-chemical (ECC) redox cycling that involves the product] and an enzyme substrate (for slow side reactions and ECC redox cycling that involve the substrate) has been developed to obtain a low detection limit for E. coli O157:H7 in an electrochemical ELISA that employs redox cycling. In our search for an alkaline phosphatase substrate/product couple that is better than the most common couple of 4-aminophenyl phosphate (APP)/4-aminophenol (AP), we compared five couples: APP/AP, hydroquinone diphosphate (HQDP)/hydroquinone (HQ), L-ascorbic acid 2-phosphate/L-ascorbic acid, 4-amino-1-naphthyl phosphate/4-amino-1-naphthol, and 1-naphthyl phosphate/1-naphthol. In particular, we examined signal-to-background ratios in ECC redox cycling using Ru(NH(3))(6)(3+) and tris(2-carboxyethyl)phosphine as an oxidant and a reductant, respectively. The ECC redox cycling that involves HQ is faster than the cycling that involves AP, whereas the side reactions and ECC redox cycling that involve HQDP are negligible compared to the APP case. These results seem to be due to the fact that the formal potential of HQ is lower than that of AP and that the formal potential of HQDP is higher than that of APP. Enzymatic amplification plus ECC redox cycling based on a HQDP/HQ couple allows us to detect E. coli O157:H7 in a wide range of concentrations from 10(3) to 10(8) colony-forming units/mL.

Facile decrease in the electron-transfer rate and surface roughness of gold by ultrasonic treatment.

This communication reports that the electron-transfer rate and surface roughness of Au electrodes can be decreased simply by ultrasonic treatment. It seems that the hydroxyl radical generated during ultrasonic treatment plays an important role, as in the case of treatment with Fenton's reagent.

Optimization of phosphatase- and redox cycling-based immunosensors and its application to ultrasensitive detection of troponin I.

The authors herein report optimized conditions for ultrasensitive phosphatase-based immunosensors (using redox cycling by a reducing agent) that can be simply prepared and readily applied to microfabricated electrodes. The optimized conditions were applied to the ultrasensitive detection of cardiac troponin I in human serum. The preparation of an immunosensing layer was based on passive adsorption of avidin (in carbonate buffer (pH 9.6)) onto indium-tin oxide (ITO) electrodes. The immunosensing layer allows very low levels of nonspecific binding of proteins. The optimum conditions for the enzymatic reaction were investigated in terms of the type of buffer solution, temperature, and concentration of MgCl(2), and the optimum conditions for antigen-antibody binding were determined in terms of incubation time, temperature, and concentration of phosphatase-conjugated IgG. Very importantly, the antigen-antibody binding at 4 °C is extremely important in obtaining reproducible results. Among the four phosphatase substrates (L-ascorbic acid 2-phosphate (AAP), 4-aminophenyl phosphate, 1-naphthyl phosphate, 4-amino-1-naphthyl phosphate) and four phosphatase products (L-ascorbic acid (AA), 4-aminophenol, 1-naphthol, 4-amino-1-naphthol), AAP and AA meet the requirements most for obtaining easy dissolution and high signal-to-background ratios. More importantly, fast AA electrooxidation at the ITO electrodes does not require modification with any electrocatalyst or electron mediator. Furthermore, tris(2-carboxyethyl)phosphine (TCEP) as a reducing agent allows fast redox cycling, along with very low anodic currents at the ITO electrodes. Under these optimized conditions, the detection limit of an immunosensor for troponin I obtained without redox cycling of AA by TCEP is ca. 100 fg/mL, and with redox cycling it is ca. 10 fg/mL. A detection limit of 10 fg/mL was also obtained even when an immunosensing layer was simply formed on a micropatterned ITO electrode. From a practical point of view, it is of great importance that ultralow detection limits can be obtained with simply prepared enzyme-based immunosensors.

Alpha-eleostearic acid suppresses proliferation of MCF-7 breast cancer cells via activation of PPARgamma and inhibition of ERK 1 / 2.

Alpha-eleostearic acid (alpha-ESA) is known to suppress the growth in cancer cells although its underlying molecular mechanisms have not been fully elucidated. The present study was designed to elucidate and evaluate the anticancer mechanism of alpha-ESA on MCF-7 breast cancer cells. Also, an attempt was made to better understand the anticancer mechanism by which alpha-ESA activated PPARgamma and attenuated the ERK1/2 MAPK phosphorylation state. The MCF-7 breast cancer cell-line and nontumorigenic MCF-10A human mammary epithelial cells were treated with alpha-ESA and compared with negative control (without treatment) and positive control groups (treated with rosiglitazone), and changes of apoptosis-related molecules, PPARgamma and pERK1/2 were examined. In MCF-7 cells treated with alpha-ESA, we found that the expression of p53, p21, and Bax was up-regulated whereas expression of Bcl-2 and procaspase-9 was down-regulated. Moreover, nuclear translocation of PPARgamma by alpha-ESA positively correlated with inhibition of ERK1/2 activation. Our data suggest that alpha-ESA can be considered to be a PPARgamma agonist and thus a candidate for a chemotherapeutic agent against breast cancer.

Fast catalytic and electrocatalytic oxidation of sodium borohydride on palladium nanoparticles and its application to ultrasensitive DNA detection.

We report an ultrasensitive DNA sensor using the rapid enhancement of electrocatalytic activity of DNA-conjugated Pd nanoparticles (NPs); the rapid enhancement results from the fast catalytic hydrolysis of NaBH(4) on Pd NPs and subsequent fast hydrogen sorption into Pd NPs.

An electrochemical immunosensor using p-aminophenol redox cycling by NADH on a self-assembled monolayer and ferrocene-modified Au electrodes.

Redox cycling of enzymatically amplified electroactive species has been widely employed for high signal amplification in electrochemical biosensors. However, gold (Au) electrodes are not generally suitable for redox cycling using a reducing (or oxidizing) agent because of the high background current caused by the redox reaction of the agent at highly electrocatalytic Au electrodes. Here we report a new redox cycling scheme, using nicotinamide adenine dinucleotide (NADH), which can be applied to Au electrodes. Importantly, p-aminophenol (AP) redox cycling by NADH is achieved in the absence of diaphorase enzyme. The Au electrodes are modified with a mixed self-assembled monolayer of mercaptododecanoic acid and mercaptoundecanol, and a partially ferrocenyl-tethered dendrimer layer. The self-assembled monolayer of long thiol molecules significantly decreases the background current of the modified Au electrodes, and the ferrocene modification facilitates easy oxidation of AP. The low amount of ferrocene on the Au electrodes minimizes ferrocene-mediated oxidation of NADH. In sandwich-type electrochemical immunosensors for mouse immunoglobulin G (IgG), an alkaline phosphatase label converts p-aminophenylphosphate (APP) into electroactive AP. The amplified AP is oxidized to p-quinoneimine (QI) by electrochemically generated ferrocenium ion. NADH reduces QI back to AP, which can be re-oxidized. This redox cycling enables a low detection limit for mouse IgG (1 pg mL(-1)) to be obtained.

Nanocatalyst-based assay using DNA-conjugated Au nanoparticles for electrochemical DNA detection.

Compared to enzymes, Au nanocatalysts show better long-term stability and are more easily prepared. Au nanoparticles (AuNPs) are used as catalytic labels to achieve ultrasensitive DNA detection via fast catalytic reactions. In addition, magnetic beads (MBs) are employed to permit low nonspecific binding of DNA-conjugated AuNPs and to minimize the electrocatalytic current of AuNPs as well as to take advantage of easy magnetic separation. In a sandwich-type electrochemical sensor, capture-probe-conjugated MBs and an indium-tin oxide electrode modified with a partially ferrocene-modified dendrimer act as the target-binding surface and the signal-generating surface, respectively. A thiolated detection-probe-conjugated AuNP exhibits a high level of unblocked active sites and permits the easy access of p-nitrophenol and NaBH 4 to these sites. Electroactive p-aminophenol is generated at these sites and is then electrooxidized to p-quinoneimine at the electrode. The p-aminophenol redox cycling by NaBH 4 offers large signal amplification. The nonspecific binding of detection-probe-conjugated AuNPs is lowered by washing DNA-linked MB-AuNP assemblies with a formamide-containing solution, and the electrocatalytic oxidation of NaBH 4 by AuNPs is minimized because long-range electron transfer between the electrode and the AuNPs bound to MBs is not feasible. The high signal amplification and low background current enable the detection of 1 fM target DNA.

Rational design of amphiphilic polymers to make carbon nanotubes water-dispersible, anti-biofouling, and functionalizable.

We report rational design of amphiphilic polymers capable of making carbon nanotubes (CNTs) highly water dispersible and resistant to biofouling; such CNTs can be conjugated with bioactive molecules so as to be potential drug delivery vehicles.

Electrochemical immunosensor using p-aminophenol redox cycling by hydrazine combined with a low background current.

Signal amplification and noise reduction are crucial for obtaining low detection limits in biosensors. Here, we present an electrochemical immunosensor in which the signal amplification is achieved using p-aminophenol (AP) redox cycling by hydrazine, and the noise level is reduced by implementing a low background current. The redox cycling is obtained in a simple one-electrode, one-enzyme format. In a sandwich-type heterogeneous immunosensor for mouse IgG, an alkaline phosphatase label converts p-aminophenyl phosphate into AP for 10 min. This generated AP is electrooxidized at an indium tin oxide (ITO) electrode modified with a partially ferrocenyl-tethered dendrimer (Fc-D). The oxidized product, p-quinone imine (QI), is reduced back to AP by hydrazine, and then AP is electrooxidized again to QI, resulting in redox cycling. Moreover, hydrazine protects AP from oxidation by air, enabling long incubation times. The small amount of ferrocene in a 0.5% Fc-D-modified ITO electrode, where 0.5% represents the ratio of ferrocene groups to dendrimer amines, results in a low background current, and this electrode exhibits high electron-mediating activity for AP oxidation. Moreover, there is insignificant hydrazine electrooxidation on this electrode, which also results in a low background current. The detection limit of the immunosensor using a 0.5% Fc-D-modified electrode is 2 orders of magnitude lower than that of a 20% Fc-D-modified electrode (10 pg/mL vs 1 ng/mL). Furthermore, the presence of hydrazine reduces the detection limit by an additional 2 orders of magnitude (100 fg/mL vs 10 pg/mL). These results indicate that the occurrence of redox cycling combined with a low background current yields an electrochemical immunosensor with a very low detection limit (100 fg/mL). Mouse IgG could be detected at concentrations ranging from 100 fg/mL to 100 microg/mL (i.e., 9 orders of magnitude) in a single assay.