High energy-charged cell factory for heterologous protein synthesis

Overexpression of gluconeogenic phosphoenolpyruvate carboxykinase (PCK) under glycolytic conditions enables Escherichia coli to maintain a greater intracellular ATP concentration and, consequently, to up-regulate genes for amino acid and nucleotide biosynthesis. To investigate the effect of a high intracellular ATP concentration on heterologous protein synthesis, we studied the expression of a foreign gene product, enhanced green fluorescence protein (eGFP), under control of the T7 promoter in E. coli BL21(DE3) strain overexpressing PCK. This strain was able to maintain twice as much intracellular ATP and to express two times more foreign protein than the control strain. These results indicate that a high energy-charged cell can be beneficial as a protein-synthesizing cell factory. The potential uses of such a cell factory are discussed

Mechanics of Graphene and CNT-polystyrene nanocomposites

We performed molecular dynamics simulation to investigate the mechanics of graphene and CNT-polystyrene nanocomposites. First, we studied the different mechanical properties of polystyrene and matched with experimental data. Then, we have investigated the interfacial and adhesive strength of graphene and CNT with polystyrene. For graphene, we considered the effect of hydrogen functionalization and different defects: stone-wales, vacancy. For CNT, we considered different chirality. Our simulation supported with analytical model give comprehensive insight into mechanism of graphene and CNT-nanocomposite

Bedeutung von Palladin für die trainingsinduzierte Anpassung der Skelettmuskulatur

In dieser Arbeit sollte erstmals geklärt werden, welchen Einfluss das Fehlen des Palladin- Proteins auf die trainingsbedingte Anpassung der Muskelzellen ausübt. Die Erkenntnisse der hier vorliegenden Arbeit bestehen darin, dass ein Fehlen der Palladin 140 und 200 kDa-Isoform nicht zu einer wesentlichen Beeinträchtigung der Muskelfaseranpassung auf Laufbandbelastungen im Skelettmuskel führt. Es konnte beobachtet werden, dass ein vierwöchiges Training gewöhnlich eine Vergrößerung der Muskelfaser im M. gastrocnemius und im M. vastus lateralis bei den KO Mäusen bewirkt. Dabei zeigten sich eine erhöhte Anzahl von zentralen Muskelzellkernen und eine unveränderte Kapillarzahl in Reaktion auf die Zunahme der Muskelfasergröße unter Trainingsbelastung. Diese morphologischen Ergebnisse konnten auch durch die Aktivität der intrazellulären Signaltransduktion, vor allem von p-ERK, bestätigt werden. In allen Muskeln der KO-Mäuse kam es zu einer Aktivitätszunahme der p-ERK, die bisher als wesentliches Merkmal der Hypertrophie im Skelettmuskel beschrieben wurde. Diese Befunde könnten auch die Grundlage für das Verständnis der Rolle des Palladin-Proteins bei der trainingsbedingten Adaptation des Skelettmuskels liefern. Zusammengefasst lässt sich vermuten, dass bei Defizienz des Palladins der Skelettmuskel durch Kompensation mit anderen Aktin-assoziierten Proteinen in der Lage ist, sich dem körperlichen Training effektiv anzupassen. Zukünftige Untersuchungen sollten einen Hinweis darauf geben, welche Mechanismen die Interaktion anderer Aktin-assoziierten FA-Proteine im Skelettmuskel als molekulare Kompensation für das Fehlen des Palladin-Proteins regulieren. Abstract For peginesatide, the first representative of a new generation of erythropoiesis-stimulating agents, a high misuse potential in elite sports is expected. Thus, the aim of the present thesis was the development of specific and sensitive mass spectrometry-based detection methods in various biological matrices relevant to doping analysis. In the context of this background, the molecular structure of the active ingredient was identified before it was synthesised and characterised by means of mass spectrometric as well as gel electrophoretic techniques. As a mass spectrometric analysis of the intact analyte is not applicable for a specific detection approach in sports drug testing, the pegylated peptide was subjected to proteolytic digestion by the serine protease subtilisin. The thereby generated pentapeptide fragment allows for a sensitive detection by means of liquid chromatography coupled to tandem mass spectrometry and fulfils the required sequence coverage. Furthermore, its xenobiotic origin is supported by a non-natural amino acid present in the fragment. Based on these results, detection assays for peginesatide in human blood (serum, plasma or dried blood spots) and urine as well as in horse serum were developed and validated in accordance with international guidelines. Proof-of-concept for the applicability of the methods was demonstrated by analysing dried blood spots, urine, and plasma samples from an in vivo study in rats, collected after a single therapeutic dose of peginesatide over a period of up to four days. In all three matrices, the characteristic pentapeptide fragment, obtained after proteolytic digestion, was unambiguously detected until the endpoint of the study, 72 h for dried blood spots and 96 h for urine and plasma specimens, respectively. Thus, within the scope of preventive doping research, specific, sensitive, and valid detection methods are applicable to routine sports drug testing since the time of drug approval (March 2012). Moreover, they are readily transferable to other doping control laboratories and allow for the detection of an illicit application of peginesatide in different biological matrices relevant to doping analysis for at least several days

Automated Brittle Fracture Rate Estimator for Steel Property Evaluation Using Deep Learning After Drop-Weight Tear Test

This study proposes an automated brittle fracture rate (BFR) estimator using deep learning. As the demand for line-pipes increases in various industries, the need for BFR estimation through dropweight tear test (DWTT) increases to evaluate steel's property. Conventional BFR or ductile fracture rate (DFR) estimation methods require an expensive 3D scanner. Alternatively, a rule-based approach is used with a single charge-coupled device (CCD) camera. However, it is sensitive to the hyper-parameter. To solve these problems, we propose an approach based on deep learning that has recently been successful in the fields of computer vision and image processing. The method proposed in this study is the first to use deep learning approach for BFR estimation. The proposed method consists of a VGG-based U-Net (VU-Net) which is inspired by U-Net and fully convolutional network (FCN). VU-Net includes a deep encoder and a decoder. The encoder is adopted from VGG19 and transferred with a pre-trained model with ImageNet. In addition, the structure of the decoder is the same as that of the encoder, and the decoder uses the feature maps of the encoder through concatenation operation to compensate for the reduced spatial information. To analyze the proposed VU-Net, we experimented with different depths of networks and various transfer learning approaches. In terms of accuracy used in real industrial application, we compared the proposed VU-Net with U-Net and FCN to evaluate the performance. The experiments showed that VU-Net was the accuracy of approximately 94.9 %, and was better than the other two, which had the accuracies of about 91.8 % and 93.7 %, respectively.11Ysciescopu

Treatment of non-odontogenic orofacial pain using botulinum toxin-A: a retrospective case series study

Background The purpose of this study was to evaluate the clinical outcomes of treatment of non-odontogenic atypical orofacial pain using botulinum toxin-A. Methods This study involved seven patients (seven females, mean age 65.1 years) who had non-odontogenic orofacial pain (neuropathic pain and atypical orofacial pain) and visited the Seoul National University Bundang Hospital between 2015 and 2017. All medication therapies were preceded by botulinum toxin-A injections, followed by injections in the insignificant effects of medication therapies. Five of the seven patients received intraoral injections in the gingival vestibule or mucosa, while the remaining two received extraoral injections in the masseter and temporal muscle areas. Results In five of the seven patients, pain after botulinum toxin-A injection was significantly reduced. Most of the patients who underwent surgery for dental implantation or facial nerve reconstruction recovered after injections. However, the pain did not disappear in two patients who reported experiencing persistent pain without any cause. Conclusions The use of botulinum toxin-A for the treatment of non-odontogenic neuropathic orofacial pain is clinically useful. It is more effective to administer botulinum toxin-A in combination with other medications and physical therapy to improve pain

A Case of Rhabdomyolysis Associated with Use of a Pneumatic Tourniquet during Arthroscopic Knee Surgery

The common causes of rhabdomyolysis include trauma, hypoxia, drugs, toxins, infections and hyperthermia. Operative insults, including direct trauma and ischemia, have the potential to cause the development of rhabdomyolysis. Pneumatic tourniquets used during arthroscopic knee surgery to prevent blood loss have led to many complications such as nerve paralysis and vascular injuries. Rhabdomyolysis can also be caused by prolonged pneumatic tourniquet application without a midapplication release, and also from an increased application pressure, but the actual incidence of this is low. In order to prevent rhabdomyolysis, the clinicians must be aware of such risks and follow strict guidelines for the application time, the midapplication release and also the inflation pressure. Vigorous hydration and postoperative patient surveillance are helpful to prevent rhabdomyolysis. We have recently experienced a case of rhabdomyolysis after the arthroscopic knee surgery, and the rhabdomyolysis could have been associated with the use of a pneumatic tourniquet