Enzymatic acyl modification of phosphatidylcholine using immobilized lipase and phospholipase A2

Ethanol‐soluble (ES) lecithin mainly contains phosphatidylcholine (PC). The incorporation of caprylic acid into PC using immobilized phospholipase A₂ (PLA₂) and lipase was investigated. The Rhizomucor meihei lipase and the porcine pancreatic PLA₂ were immobilized on the hydrophobic resin Diaion HP‐20 and the modification was carried out in hexane as solvent. HPTLC with densitometer technique was successfully used for monitoring the production of structured phospholipids (PL) (ML‐type PC, MM‐type PC, and lysophosphatidylcholine; L: long‐chain fatty acid, M: medium‐chain fatty acid). The various parameters such as the effects of reaction temperature, enzyme loading, and the effect of molar proportion of substrate were studied in order to determine the optimum reaction conditions for the acidolysis reaction. The optimal operating conditions for the PLA₂‐catalyzed reaction were obtained as 50°C temperature, 50% (wt/wt of substrate) enzyme loading, and a 1:12 molar proportion of PC/caprylic acid. For the lipase‐catalyzed reaction, the optimized temperature was the same as for PLA₂, but the enzyme loading and molar proportion were slightly lower, i.e., 40 % w/w of substrate and 1:9 PC/caprylic acid, respectively. The effects of these parameters on the production of structured PL were compared. Under these optimal conditions, the ML‐type PC content was higher in the PLA₂‐catalyzed reaction, i.e., 45.29 mol%, and in the lipase‐catalyzed reaction it was 38.74 mol%.     

Effect of Secondary Structure and Side Chain Length of Hydrophobic Amino Acid Residues on the Antimicrobial Activity and Toxicity of 14-Residue-Long de novo AMPs

Herein we report the efficacy and toxicity of three de novo designed cationic antimicrobial peptides (AMPs) LL-14, VV-14 and ββ-14, where side chains of the hydrophobic amino acids were reduced gradually. The AMPs showed broad-spectrum antimicrobial activity against three pathogens from the ESKAPE group and two fungal strains. This study showed that side chains which are either too long or too short increase toxicity and lower antimicrobial activity, respectively. VV-14 was found to be non-cytotoxic and highly potent under physiological salt concentrations against several pathogens, especially Salmonella typhi TY2. These AMPs acted via membrane deformation, depolarization, and lysis. The activity of the AMPs is related to their ability to take on amphipathic helical conformations in the presence of microbial membrane mimics. Among AMPs with the same charge, hydrophobic interactions between the side chains of the residues with cell membrane lipids determine their antimicrobial potency and cytotoxicity. Strikingly, an optimum hydrophobic interaction is the crux of generating highly potent non-cytotoxic AMPs.     

Design and analysis of a novel mechanical loading machine for dynamic in vivo axial loading

This paper describes the construction of a loading machine for performing in vivo, dynamic mechanical loading of the rodent forearm. The loading machine utilizes a unique type of electromagnetic actuator with no mechanically resistive components (servotube), allowing highly accurate loads to be created. A regression analysis of the force created by the actuator with respect to the input voltage demonstrates high linear correlation (R(2) = 1). When the linear correlation is used to create dynamic loading waveforms in the frequency (0.5-10 Hz) and load (1-50 N) range used for in vivo loading, less than 1% normalized root mean square error (NRMSE) is computed. Larger NRMSE is found at increased frequencies, with 5%-8% occurring at 40 Hz, and reasons are discussed. Amplifiers (strain gauge, linear voltage displacement transducer (LVDT), and load cell) are constructed, calibrated, and integrated, to allow well-resolved dynamic measurements to be recorded at each program cycle. Each of the amplifiers uses an active filter with cutoff frequency at the maximum in vivo loading frequencies (50 Hz) so that electronic noise generated by the servo drive and actuator are reduced. The LVDT and load cell amplifiers allow evaluation of stress-strain relationships to determine if in vivo bone damage is occurring. The strain gauge amplifier allows dynamic force to strain calibrations to occur for animals of different sex, age, and strain. Unique features are integrated into the loading system, including a weightless mode, which allows the limbs of anesthetized animals to be quickly positioned and removed. Although the device is constructed for in vivo axial bone loading, it can be used within constraints, as a general measurement instrument in a laboratory setting.     

Fibrolamellar carcinoma of the liver--an unusual presentation

Non-bacterial thrombotic endocarditis was found at autopsy in a 17 year old male patient of fibrolamellar type of hepatocellular carcinoma with pericardial metastases. This had resulted in multiple embolic cerebral infarcts with long standing hemiplegia and later death due to acute left ventricular failure.     

A semi-automated measurement technique for the assessment of radiolucency

The assessment of radiolucency around an implant is qualitative, poorly defined and has low agreement between clinicians. Accurate and repeatable assessment of radiolucency is essential to prevent misdiagnosis, minimize cases of unnecessary revision, and to correctly monitor and treat patients at risk of loosening and implant failure. The purpose of this study was to examine whether a semi-automated imaging algorithm could improve repeatability and enable quantitative assessment of radiolucency. Six surgeons assessed 38 radiographs of knees after unicompartmental knee arthroplasty for radiolucency, and results were compared with assessments made by the semi-automated program. Large variation was found between the surgeon results, with total agreement in only 9.4% of zones and a kappa value of 0.602; whereas the automated program had total agreement in 81.6% of zones and a kappa value of 0.802. The software had a ‘fair to excellent’ prediction of the presence or the absence of radiolucency, where the area under the curve of the receiver operating characteristic curves was 0.82 on average. The software predicted radiolucency equally well for cemented and cementless implants (p = 0.996). The identification of radiolucency using an automated method is feasible and these results indicate that it could aid the definition and quantification of radiolucency.     

Outcome aware ranking in value creation networks

In this paper, we consider a natural ranking problem that arises in settings in which a community of people (or agents) are engaged in regular interactions with an end goal of creating value. Examples of such scenarios are academic collaboration networks, creative collaborations, and interactions between agents of a service delivery organization. For instance, consider a service delivery organization which essentially resolves a sequence of service requests from its customers by deploying its agents to resolve the requests. Typically, resolving a request requires interaction between multiple agents and results in an outcome (or value). The outcome could be success or failure of problem resolution or an index of customer satisfaction. For this scenario, the ranking of the agents of the network should take into account two aspects: importance of the agents in the network structure that arises as a result of interactions and the value generated by the interactions involving the respective agents. Such a ranking can be used for several purposes such as identifying influential agents of the interaction network, effective and efficient spreading of messages in the network. In this paper, we formally model the above ranking problem and develop a novel algorithm for computing the ranking. The key aspect of our approach is creating special nodes in the interaction network corresponding to the outcomes and endowing them independent, external status. The algorithm then iteratively spreads the external status of the outcomes to the agents based on their interactions and the outcome of those interactions. This results in an eigenvector like formulation, which results in a method requiring computing the inverse of a matrix rather than the eigenvector. We present several theoretical characterizations of our algorithmic approach. We present experimental results on the public domain real-life datasets from the Internet Movie Database and a dataset constructed by retrieving impact and citation ratings for papers listed in the DBLP database.     

Preferential cell response to anisotropic electro-spun fibrous scaffolds under tension-free conditions

Anisotropic alignment of collagen fibres in musculoskeletal tissues is responsible for the resistance to mechanical loading, whilst in cornea is responsible for transparency. Herein, we evaluated the response of tenocytes, osteoblasts and corneal fibroblasts to the topographies created through electro-spinning and solvent casting. We also evaluated the influence of topography on mechanical properties. At day 14, human osteoblasts seeded on aligned orientated electro-spun mats exhibited the lowest metabolic activity (P < 0.001). At day 5 and at day 7, no significant difference was observed in metabolic activity of human corneal fibroblasts and bovine tenocytes respectively seeded on different scaffold conformations (P > 0.05). Osteoblasts and corneal fibroblasts aligned parallel to the direction of the aligned orientated electro-spun mats, whilst tenocytes aligned perpendicular to the aligned orientated electro-spun mats. Mechanical evaluation demonstrated that aligned orientated electro-spun fibres exhibited significant higher stress at break values than their random aligned counterparts (P < 0.006) and random orientated electro-spun fibres exhibited significant higher strain at break values than the aligned orientated scaffolds (P < 0.006). While maintaining fibre structure, we also developed a co-deposition method of spraying and electro-spinning, which enables the incorporation of microspheres within the three-dimensional structure of the scaffold.     

Gas hold-up behavior of mechanically agitated gas-liquid reactors using pitched blade downflow turbines

Fractional gas holdup was measured in 0.57, 1.0, and 1.5 m i.d. vessels. Pitched blade downflow turbines (PTD) were used as the impeller. Design details of the impeller, such as the impeller diameter (0.22 T to 0.5 T) and blade width (0.25 D to 0.4 D), were studied. The effect of sparger type, geometry and size on fractional gas hold-up has been investigated in detail. Four different types of spargers (pipe, conical, ring and concentric ring spargers) were used. Sparger location was varied for all the types studied. Further, design details of the ring sparger, which gave the highest hold-up were then studied in detail. These included ring diameter, number of holes and hole size. All the reported correlations for fractional gas hold-up in mechanically agitated gas-liquid reactors were tested and compared. A better correlation has been developed for pitched blade turbines.