Characterizations and Inkjet Printing of Carbon Black Electrodes for Dielectric Elastomer Actuators.

Dielectric elastomer actuators (DEAs) have been proposed as a promising technology for developing soft robotics and stretchable electronics due to their large actuation. Among available fabrication techniques, inkjet printing is a digital, mask-free, material-saving, and fast technology, making it versatile and appealing for fabricating DEA electrodes. However, there is still a lack of suitable materials for inkjet-printed electrodes. In this study, multiple carbon black (CB) inks were developed and tested as DEA electrodes inkjet-printed on acrylic membranes (VHB). Triethylene glycol monomethyl ether (TGME) and chlorobenzene (CLB) were selected to disperse CB. The inks' stability, particle size, surface tension, viscosity, electrical resistance, and printability were characterized. The DEA with Ink-TGME/CLB (mixture solvent) electrodes obtained 80.63% area strain, a new benchmark for the DEA actuation with CB powder electrodes on VHB. The novelty of this work involves the disclosure of a new ink recipe (TGME/CLB/CB) for inkjet printing that can obtain stable drop formations with a small nozzle (17 × 17 µm), high resolution (∼25 µm, approaching the limit of drop-on-demand inkjet printing), and the largest area strain of DEAs under similar conditions, distinguishing this contribution from the previous works, which is important for the fabrication and miniaturization of DEA-based soft and stretchable electronics.

One Soft Step: Bio-Inspired Artificial Muscle Mechanisms for Space Applications.

Soft robots, devices with deformable bodies and powered by soft actuators, may fill a hitherto unexplored niche in outer space. All space-bound payloads are heavily limited in terms of mass and volume, due to the cost of launch and the size of spacecraft. Being constructed from stretchable materials allows many possibilities for compacting soft robots for launch and later deploying into a much larger volume, through folding, rolling, and inflation. This morphability can also be beneficial for adapting to operation in different environments, providing versatility, and robustness. To be truly soft, a robot must be powered by soft actuators. Dielectric elastomer transducers (DETs) offer many advantages as artificial muscles. They are lightweight, have a high work density, and are capable of artificial proprioception. Taking inspiration from nature, in particular the starfish podia, we present here bio-inspired inflatable DET actuators powering low-mass robots capable of performing complex motion that can be compacted to a fraction of their operating size.

Modelling dielectric elastomer circuit networks for soft biomimetics.

In order to obtain entirely soft bio-inspired robots, fully soft electronic circuits are needed. Dielectric elastomers (DEs) are electroactive polymers that have demonstrated multifunctionality. The same material can achieve different tasks like actuation, sensing, or energy harvesting. It has been shown that basic logic and memory functions can be realized with similar soft structures by combining multiple DE actuators and DE switches. Thus it would be possible to build, with the same materials and processes, a soft structure that mimics a biological being with all these capabilities. This contribution is focused on the modelling of the aforementioned soft electro-mechanical circuit networks. It is here reported the building process of a comprehensive SIMULINK model including the electro-mechanical behaviour of DE logic units and their interconnections. Conventional models deal with a single aspect of DEs, generating complex finite-element simulations. This contribution, based on a former model for an inverter-based DEO, shows how to integrate these various mathematical models and, with the help of direct measurements, create a software representation of DE circuit networks. This work is intended to demonstrate the validity of a recently introduced model and apply it to more complex circuit networks that have a higher number of components. Since, at the present state, the building processes are by hand, adding components generates more variability due to sample-to-sample variation and human error. Despite this, the model still shows a qualitatively good prediction of the devices' behaviour. Furthermore, the introduction of new materials and automatic processes will help to reduce this variability, allowing the model to reach even better performance.

Dielectric Elastomer Actuator Driven Soft Robotic Structures With Bioinspired Skeletal and Muscular Reinforcement.

Natural motion types found in skeletal and muscular systems of vertebrate animals inspire researchers to transfer this ability into engineered motion, which is highly desired in robotic systems. Dielectric elastomer actuators (DEAs) have shown promising capabilities as artificial muscles for driving such structures, as they are soft, lightweight, and can generate large strokes. For maximum performance, dielectric elastomer membranes need to be sufficiently pre-stretched. This fact is challenging, because it is difficult to integrate pre-stretched membranes into entirely soft systems, since the stored strain energy can significantly deform soft elements. Here, we present a soft robotic structure, possessing a bioinspired skeleton integrated into a soft body element, driven by an antagonistic pair of DEA artificial muscles, that enable the robot bending. In its equilibrium state, the setup maintains optimum isotropic pre-stretch. The robot itself has a length of 60 mm and is based on a flexible silicone body, possessing embedded transverse 3D printed struts. These rigid bone-like elements lead to an anisotropic bending stiffness, which only allows bending in one plane while maintaining the DEA's necessary pre-stretch in the other planes. The bones, therefore, define the degrees of freedom and stabilize the system. The DEAs are manufactured by aerosol deposition of a carbon-silicone-composite ink onto a stretchable membrane that is heat cured. Afterwards, the actuators are bonded to the top and bottom of the silicone body. The robotic structure shows large and defined bimorph bending curvature and operates in static as well as dynamic motion. Our experiments describe the influence of membrane pre-stretch and varied stiffness of the silicone body on the static and dynamic bending displacement, resonance frequencies and blocking forces. We also present an analytical model based on the Classical Laminate Theory for the identification of the main influencing parameters. Due to the simple design and processing, our new concept of a bioinspired DEA based robotic structure, with skeletal and muscular reinforcement, offers a wide range of robotic application.

Modeling of dielectric elastomer oscillators for soft biomimetic applications.

Biomimetic, entirely soft robots with animal-like behavior and integrated artificial nervous systems will open up totally new perspectives and applications. However, until now, most presented studies on soft robots were limited to only partly soft designs, since all solutions at least needed conventional, stiff electronics to sense, process signals and activate actuators. We present a novel approach for a set up and the experimental validation of an artificial pace maker that is able to drive basic robotic structures and act as artificial central pattern generator. The structure is based on multi-functional dielectric elastomers (DEs). DE actuators, DE switches and DE resistors are combined to create complex DE oscillators (DEOs). Supplied with only one external DC voltage, the DEO autonomously generates oscillating signals that can be used to clock a robotic structure, control the cyclic motion of artificial muscles in bionic robots or make a whole robotic structure move. We present the basic functionality, derive a mathematical model for predicting the generated signal waveform and verify the model experimentally.

Soft Dielectric Elastomer Oscillators Driving Bioinspired Robots.

Entirely soft robots with animal-like behavior and integrated artificial nervous systems will open up totally new perspectives and applications. To produce them, we must integrate control and actuation in the same soft structure. Soft actuators (e.g., pneumatic and hydraulic) exist but electronics are hard and stiff and remotely located. We present novel soft, electronics-free dielectric elastomer oscillators, which are able to drive bioinspired robots. As a demonstrator, we present a robot that mimics the crawling motion of the caterpillar, with an integrated artificial nervous system, soft actuators and without any conventional stiff electronic parts. Supplied with an external DC voltage, the robot autonomously generates all signals that are necessary to drive its dielectric elastomer actuators, and it translates an in-plane electromechanical oscillation into a crawling locomotion movement. Therefore, all functional and supporting parts are made of polymer materials and carbon. Besides the basic design of this first electronic-free, biomimetic robot, we present prospects to control the general behavior of such robots. The absence of conventional stiff electronics and the exclusive use of polymeric materials will provide a large step toward real animal-like robots, compliant human machine interfaces, and a new class of distributed, neuron-like internal control for robotic systems.

VEGF-ablation therapy reduces drug delivery and therapeutic response in ECM-dense tumors.

The inadequate transport of drugs into the tumor tissue caused by its abnormal vasculature is a major obstacle to the treatment of cancer. Anti-vascular endothelial growth factor (anti-VEGF) drugs can cause phenotypic alteration and maturation of the tumor's vasculature. However, whether this consistently improves delivery and subsequent response to therapy is still controversial. Clinical results indicate that not all patients benefit from antiangiogenic treatment, necessitating the development of criteria to predict the effect of these agents in individual tumors. We demonstrate that, in anti-VEGF-refractory murine tumors, vascular changes after VEGF ablation result in reduced delivery leading to therapeutic failure. In these tumors, the impaired response after anti-VEGF treatment is directly linked to strong deposition of fibrillar extracellular matrix (ECM) components and high expression of lysyl oxidases. The resulting condensed, highly crosslinked ECM impeded drug permeation, protecting tumor cells from exposure to small-molecule drugs. The reduced vascular density after anti-VEGF treatment further decreased delivery in these tumors, an effect not compensated by the improved vessel quality. Pharmacological inhibition of lysyl oxidases improved drug delivery in various tumor models and reversed the negative effect of VEGF ablation on drug delivery and therapeutic response in anti-VEGF-resistant tumors. In conclusion, the vascular changes after anti-VEGF therapy can have a context-dependent negative impact on overall therapeutic efficacy. A determining factor is the tumor ECM, which strongly influences the effect of anti-VEGF therapy. Our results reveal the prospect to revert a possible negative effect and to potentiate responsiveness to antiangiogenic therapy by concomitantly targeting ECM-modifying enzymes.

Screening of commercial hydrolases for the degradation of ochratoxin A.

Ochratoxin A (OTA) is a nephrotoxic and carcinogenic mycotoxin. The toxin is a common contaminant of various foods and feeds and poses a serious threat to the health of both humans and animals. A number of commercial hydrolases were screened for the ability to degrade OTA to nontoxic compounds. A crude lipase from Aspergillus niger (Amano A) proved to substantially hydrolyze OTA to the nontoxic OTalpha and phenylalanine, as confirmed by HPLC with fluorescence detection. The enzyme was purified by anion exchange chromatography to homogeneity. Activity staining of the purified enzyme with alpha-naphthyl acetate/Fast Red revealed only one band exhibiting hydrolytic activity. The specific activity of the purified enzyme toward OTA was 2.32 units/mg.

Animal studies on the in vivo stability of 211At-labelled albumin particles.

The alpha-emitter astatine-211 appears as one of the most suitable representatives for radiotherapy owing to its excellent decay properties. We report the studies in mice on the stability of the astatine bond to HSA-particles. Microspheres stick fast in the lung capillaries. There is a comparatively very slow decrease of radioactivity in the lungs. The 211At-labelled microspheres appear quite stable in vivo compared with a series of other biomolecules and so give a potential possibility for using the excellent radiophysical features of 211At in endogenous therapy of malignant tumours.

Induction of complete regressions of oncogene-induced breast tumors in mice.

Over the past decade, mouse models of cancer have come to resemble human disease much more closely than simple subcutaneous or orthotopic systems. Intervention strategies that work on these new model systems are more likely to have an impact clinically. We have shown recently that antiangiogenic stress imposed by loss of Id protein in endothelial progenitor cells results in dramatic central necrosis in breast tumors initiated in mice by overexpression of the her2/neu oncogene. Tumor cells remain viable at the periphery, perhaps via the hypoxic response pathway which allows the lesions to expand. Inhibition of this pathway by the inactivation of the Hif-1alpha chaperone Hsp90 in combination with antiangiogenic stress leads to the first reported complete regression of these aggressive breast tumors.

Directed evolution of an esterase from Pseudomonas fluorescens. Random mutagenesis by error-prone PCR or a mutator strain and identification of mutants showing enhanced enantioselectivity by a resorufin-based fluorescence assay.

The gene encoding an esterase from Pseudomonas fluorescens (PFE) was subjected to random mutagenesis by error-prone PCR or by using the mutator strain Epicurian coli XL1-Red. Enzyme libraries were then created in microtiter plates by expression of PFE-variants in E. coli. These were assayed for improved enantioselectivity in a Beckman robot system using optically pure (R)- or (S)-3-phenylbutyric acid resorufin esters, resulting in the identification of several mutants showing up to almost two-fold enantioselectivity (E(true) = 5.2 to 6.6) compared to wild-type PFE (E(true) = 3.5).

Esterases from Bacillus subtilis and B. stearothermophilus share high sequence homology but differ substantially in their properties.

A novel esterase from Bacillus subtilis (BsubE) was cloned, functionally expressed in Escherichia coli and biochemically characterized. BsubE shows high homology (74% identity, >95% homology) to an esterase from the thermophilic B. stearothermophilus (BsteE). Both enzymes were efficiently expressed in E. coli, using a L-rhamnose-expression system [11,500 units/l (BsteE), 3,400 units/l (BsubE)] and were purified by Ni-nitrilotriacetic acid chromatography, yielding specific activities of 70 units/mg (BsteE) and 40 units/mg (BsubE), as determined by the hydrolysis of p-nitrophenyl acetate. Despite the high homology, both esterases revealed remarkable differences in their properties. As expected, the esterase from the thermophilic organism showed significantly higher temperature stability. Whereas BsteE showed highest activity at 65-70 degrees C, BsubE was almost inactivated at 50 degrees C. Moreover, both enzymes showed quite different substrate patterns in the hydrolysis of various esters. Whilst the B. subtilis esterase accepted esters with a branched alcohol moiety well, the B. stearothermophilus esterase was more useful in the hydrolysis of substrates with a sterically demanding carboxylic acid group. BsteE showed excellent enantioselectivity ( E>100) in the kinetic resolution of menthyl acetate and even accepted the bulky menthyl benzoate as substrate ( E=19). In contrast, BsubE converted 1-phenethylacetate with higher selectivity ( E>150 vs E=8).

Familial occurrence of accessory atrioventricular pathways (preexcitation syndrome).

Accessory atrioventricular pathways, the anatomical structures responsible for the preexcitation syndromes, may result from a developmental failure to eradicate the remnants of the atrioventricular connections present during cardiogenesis. To study whether preexcitation syndromes could also be transmitted genetically, we determined the prevalence of preexcitation in the first-degree relatives of 383 of 456 consecutive patients (84 percent) with electrophysiologically proved accessory pathways. We compared the observed prevalence of preexcitation among the 2343 first-degree relatives with the frequency reported in the general population (0.15 percent). For 13 of the 383 index patients (3.4 percent), accessory pathways were documented in one or more first-degree relatives. At least 13 of the 2343 relatives identified (0.55 percent) had preexcitation; this prevalence was significantly higher than that in the general population (P less than 0.0001). Identification of affected relatives may have been incomplete because clinical information was obtained only about symptomatic relatives. Patients with familial preexcitation have a higher incidence of multiple accessory pathways and possibly an increased risk of sudden cardiac death. Our data suggest a hereditary contribution to the development of accessory pathways in humans. The pattern of inheritance appears to be autosomal dominant.

Induction of thyroid neoplasms in the rat. A histochemical, immunohistochemical, autoradiographic and ultrastructural study.

Papillary carcinomas of the thyroid gland induced in rats by combined methylthiouracil (MTU)/nitrosomethylurea treatment were studied electron microscopically and histochemically for their acid phosphatase activity, radioactive iodine uptake and for their thyroglobulin synthesis. The papillary carcinomas accumulated radioactive iodine, synthesized thyroglobulin and showed a perinuclear localization of acid phosphatase activity in the cytoplasm. After omitting the reactive thyrotropic effect (cessation of MTU administration) a decrease of both iodine organification and acid phosphatase activity was observed. The synthesis of thyroglobulin was also reduced. The changes in the ultrastructure of tumour cells corresponded to the respective functional activities. The results suggest that the papillary carcinomas may have a functional dependence on thyroid-stimulating hormone.

A 38 base pair insertion in the pro alpha 2(I) collagen gene of a patient with Marfan syndrome.

Abnormalities in type I collagen have been recognized in a number of connective tissue disorders. In the Marfan syndrome, an autosomal dominant condition producing a generalized abnormality in connective tissue, no consistent abnormality has been identified, although one individual has been found to have an elongated pro alpha 2(I) collagen chain [Byers et al, Proc Natl Acad Sci USA 78:7745, 1981]. To determine the nature of the alteration in the gene that produced this abnormality, we studied the pro alpha 2(I) gene from this individual by genomic blotting and gene cloning. Genomic mapping studies detected no abnormalities. However, analysis of the cloned segment of the pro alpha 2(I) collagen gene from the Marfan individual indicates that the gene contains a 38 base pair insertion in an intron near the collagenase cleavage site. Although the relationship of this insertion to the protein abnormality is unclear, it may be a useful marker for the diagnosis of the Marfan syndrome.

[Evaluation of intratumoral bleomycin application in a tumor model and in a clinical pilot study]. / Beurteilung der intratumoralen (i.t.) Bleomycin (Bleo)- Applikation am Tumor-Modell und in der klinischen Pilotstudie.

In a total of 21 patients with squamous cell carcinomas of the oral mucosa 26.9% of the total volume of 57Co-Bleomycin injected into the tumor was found in the blood as early as 10 min following injection. Examinations using a gamma camera demonstrated that 94.6% of the substance have cleared the tumor tissue after just 1 h. This sharp and rapid drop in activity, which was particularly pronounced in carcinomas of the tongue and the floor of the mouth, might be caused by rapid resorption due to the high degree of vascularization of the maxillofacial region. Autoradiography showed deposits of the substance in tumor-free areas of the operation specimens. It seems that the postulated advantages of intratumoral application--increased concentration and depot effect in the tumor tissue--are no longer tenable, thus large-scale clinical trials with intratumoral bleomycin treatment cannot be justified.

[Experimental research following intratumor bleomycin use in the nude mouse model of oral mucosa cancer and the clinical pilot study]. / Experimentelle Untersuchungen nach intratumoraler Bleomycin-Applikation am nude-Maus-Modell des Mundschleimhautkarzinoms und klinische Pilotstudie.

Experiments concerning the kinetics and bio-distribution of 57Co-Bleomycin in three different lines of the squamous cell carcinoma of the oral cavity in a nude-mouse model yielded a more than tenfold higher concentration after a single intratumoral (i.t.) application of an aqueous solution of 57Co-labelled Bleomycin in spite of a remarkable radioactivity-wash in the tumor up to 48 hours post applicationem, compared to the intravenous (i.v.) injection. For the investigation of the radiopharmacon within the tumor we used the macro-autoradiographic method. The xenografts have been removed and cut (5 micrograms) 1;5; 24, and 48 hours after the Bleomycin administration, respectively. These preparations have been covered with an autoradiographic film and exposed for about three weeks. After this an inhomogenous distribution of the radioactive nuclide was produced within the tumor, and a particularly high activity-concentration could be demonstrated in the necrotic tumor areas. The results obtained from the animal experiments have been corroborated by a pilot-study consisting of i.t. Bleomycin application in nine patients with a carcinoma of the oral mucosa.

The influence of isotopic and nonisotopic carriers on the biodistribution and biokinetics of M3+-citrate complexes.

The influence of carrier amounts of Fe, Ga, and Tm on the biodistribution of 67Ga-, 59Fe-, and 167Tm-citrate in mice was investigated. Our results suggest that 167Tm, unlike 67Ga and 59Fe, is not transported by transferrin in the blood. Of the three radioisotopes tested, 167Tm had the highest tumor/background ratio (10 h after the injection). The application of Fe carrier led to an enhancement of the elimination of 67Ga from the blood and muscles, resulting in a better tumor/background ratio.