A facile method for isolation of recombinant human apolipoprotein A-I from E. coli

Apolipoprotein (apo) A-I is the major protein component of high-density lipoprotein (HDL) and plays key roles in the Reverse Cholesterol Transport pathway. In the past decade, reconstituted HDL (rHDL) has been employed as a therapeutic agent for treatment of atherosclerosis. The ability of rHDL to promote cholesterol efflux from peripheral cells has been documented to reduce the size of atherosclerotic plaque lesions. However, development of apoA-I rHDL-based therapeutics for human use requires a cost effective process to generate an apoA-I product that meets "Good Manufacturing Practice" standards. Methods available for production and isolation of unmodified recombinant human apoA-I at scale are cumbersome, laborious and complex. To overcome this obstacle, a streamlined two-step procedure has been devised for isolation of recombinant untagged human apoA-I from E. coli that takes advantage of its ability to re-fold to a native conformation following denaturation. Heat treatment of a sonicated E. coli supernatant fraction induced precipitation of a large proportion of host cell proteins (HCP), yielding apoA-I as the major soluble protein. Reversed-phase HPLC of this material permitted recovery of apoA-I largely free of HCP and endotoxin. Purified apoA-I possessed α-helix secondary structure, formed rHDL upon incubation with phospholipid and efficiently promoted cholesterol efflux from cholesterol loaded J774 macrophages

Perspectives of Cross-Border Security of Poland

Celem pracy jest zdefiniowanie bezpieczeństwa transgranicznego, próba wytyczenia nowego kierunku jego rozwoju, oraz ukazanie istotnych problemów dotyczących funkcjonowania systemu ochrony granicy państwowej i sformułowanie wskazówek służących ich rozwiązaniu. Materiał badawczy stanowi literatura fachowa związana z szeroko pojętym bezpieczeństwem. Metoda badawcza to analiza tekstu i danych statystycznych. Wyniki badań koncentrują się na przedstawieniu nieprawidłowości i różnic pomiędzy założeniami teoretycznymi, a stanem rzeczywistym bezpieczeństwa transgranicznego RP. Wnioski płynące z analizowanych badań pozwalają stwierdzić, iż poprawa ,,Założeń wieloletniej koncepcji funkcjonowania Straży Granicznej na lata 2009-2015» znacząco wpłynie na proces umacniania bezpieczeństwa transgranicznego Polski.The aim of this paper is to define the cross-border security and to attempt the delineation of a new direction for its development, as well as to show the important issues relating to the functioning of the state border protection and make suggestions for their solution. The research material is literature related to the wider security, as well as the Border Guard statistics. The test method is the analysis of text and statistics. The results of the research focused on the presentation of the discrepancies and differences between the theoretical assumptions and the actual state of the Polish cross-border security. The conclusions of the included studies allow us to conclude that the long-term improvement ,,Assumptions Border Guard operation concept for 2009-2015” significantly affect the process of strengthening Polish cross-border security

Perspektywy bezpieczeństwa przewozu towarów niebezpiecznych transportem drogowym w Rzeczypospolitej Polskiej

Przemiany społeczno-technologiczne wraz z postępem cywilizacyjnym pozytywnie wpływają na rozwój wielu dziedzin gospodarki, a zarazem generują szereg różnych zagrożeń, którym należy przeciwdziałać. Coraz częściej można usłyszeć o różnego rodzaju katastrofach i awariach w zakładach przemysłowych, instalacjach technologicznych, urządzeniach oraz środkach transportu. Ryzyko wystąpienia tej odmiany zagrożeń zdecydowanie wzrasta, gdy przedmiotami w procesie produkcyjnym, magazynowaniu i transporcie są towary niebezpieczne. Postęp cywilizacyjno-technologiczny, przekładający się na wzrost możliwości przemysłu sprawia, iż wzrasta popyt na usługi komunikacyjno-transportowe. Ze względu na możliwość szybkiego dostarczenia towarów w relacji door-to-door najczęściej wybieraną a zarazem najpowszechniejszą gałęzią transportu jest transport samochodowy, który zdominował rynek przewozów

Modeling Hydrodynamics and Water Quality for Wachusett Reservoir

The purpose of this paper is to present applications of long-term and multi-faceted approaches undertaken for the Division of Water Supply Protection of the Massachusetts Department of Conservation and Recreation (DCR) in modeling hydrodynamics and water quality in Wachusett Reservoir. The modeling work is part of a larger effort focused on the two reservoir (Quabbin and Wachusett) source of currently unfiltered drinking water for the Boston metropolitan area, with the long term research goal of providing a rational basis for watershed and reservoir management decisions through measurements of water quality and modeling of significant transport and transformation processes. Modeling of reservoir hydrodynamics and water quality has included extensive use of CEQUAL W2, a two dimensional (longitudinal segments, depth layers, lateral homogeneity) finite difference model available in the public domain. Assessment and calibration of annual water budgets have highlighted strengths and weaknesses of available data in properly characterizing water quantity. Wachusett, the terminal supply reservoir, can be significantly affected by transfers from the larger, more pristine, Quabbin reservoir; modeling has illuminated key impacts of this transfer. Another aspect of the project has been to assess inputs and transformations of natural organic matter (NOM) due to its influence on subsequent oxidant/disinfectant demands and by-product formation. The current modeling work is directed as assessing potential impacts of contaminant spills (sewage, hydrocarbons) into the reservoir on water quality in the overall reservoir and at the Cosgrove water supply intake. Impacts of spill date (season) and location as well as contaminant type are being investigated through use of both the two dimensional CE QUAL W2 model and a three dimensional computational fluid dynamics (CFD) model developed for the Thomas Basin. This paper will summarize and highlight key findings from the recent contaminant spill modeling work

Modeling Hydrodynamics and Water Quality for Wachusett Reservoir

The purpose of this paper is to present applications of long-term and multi-faceted approaches undertaken for the Division of Water Supply Protection of the Massachusetts Department of Conservation and Recreation (DCR) in modeling hydrodynamics and water quality in Wachusett Reservoir. The modeling work is part of a larger effort focused on the two reservoir (Quabbin and Wachusett) source of currently unfiltered drinking water for the Boston metropolitan area, with the long term research goal of providing a rational basis for watershed and reservoir management decisions through measurements of water quality and modeling of significant transport and transformation processes. Modeling of reservoir hydrodynamics and water quality has included extensive use of CEQUAL W2, a two dimensional (longitudinal segments, depth layers, lateral homogeneity) finite difference model available in the public domain. Assessment and calibration of annual water budgets have highlighted strengths and weaknesses of available data in properly characterizing water quantity. Wachusett, the terminal supply reservoir, can be significantly affected by transfers from the larger, more pristine, Quabbin reservoir; modeling has illuminated key impacts of this transfer. Another aspect of the project has been to assess inputs and transformations of natural organic matter (NOM) due to its influence on subsequent oxidant/disinfectant demands and by-product formation. The current modeling work is directed as assessing potential impacts of contaminant spills (sewage, hydrocarbons) into the reservoir on water quality in the overall reservoir and at the Cosgrove water supply intake. Impacts of spill date (season) and location as well as contaminant type are being investigated through use of both the two dimensional CE QUAL W2 model and a three dimensional computational fluid dynamics (CFD) model developed for the Thomas Basin. This paper will summarize and highlight key findings from the recent contaminant spill modeling work

Real-Time and Remote Construction Progress Monitoring with a Quadruped Robot Using Augmented Reality

Construction progress monitoring involves a set of inspection tasks with repetitive in-person observations on the site. The current manual inspection process in construction is time-consuming, inefficient and inconsistent mainly due to human limitations in the ability to persistently and accurately walkthrough the job site and observe the as-built status of which robots are considerably better. Enabling the process of visual inspection with a real-time and remote inspection capability using robots can provide more frequent and accessible construction progress data for inspectors to improve the quality of inspection and monitoring. Also, integrating remote inspection with an Augmented Reality (AR) platform can help the inspector to verify as-planned BIM data with the as-built status. This paper proposes a new approach to perform remote monitoring of the construction progress in real-time using a quadruped robot and an AR solution. The proposed computational framework in this study uses a cloud-based solution to integrate the quadruped robot’s control for remote navigation through the construction site with 360° live-stream video of the construction status, as well as a real-time AR solution to visualize and compare the as-built status with as-planned BIM geometry. The implementation of the proposed framework is discussed, and the developed framework is evaluated in two use cases through experimental investigations

Combination of oxyanion Gln114 Mutation and medium engineering to influence the enantioselectivity of thermophilic lipase from geobacillus zalihae

The substitution of the oxyanion Q114 with Met and Leu was carried out to investigate the role of Q114 in imparting enantioselectivity on T1 lipase. The mutation improved enantioselectivity in Q114M over the wild-type, while enantioselectivity in Q114L was reduced. The enantioselectivity of the thermophilic lipases, T1, Q114L and Q114M correlated better with log p as compared to the dielectric constant and dipole moment of the solvents. Enzyme activity was good in solvents with log p < 3.5, with the exception of hexane which deviated substantially. Isooctane was found to be the best solvent for the esterification of (R,S)-ibuprofen with oleyl alcohol for lipases Q114M and Q114L, to afford E values of 53.7 and 12.2, respectively. Selectivity of T1 was highest in tetradecane with E value 49.2. Solvents with low log p reduced overall lipase activity and dimethyl sulfoxide (DMSO) completely inhibited the lipases. Ester conversions, however, were still low. Molecular sieves employed as desiccant were found to adversely affect catalysis in the lipase variants, particularly in Q114M. The higher desiccant loading also increased viscosity in the reaction and further reduced the efficiency of the lipase-catalyzed esterifications

Real-Time and Remote Construction Progress Monitoring with a Quadruped Robot Using Augmented Reality

Construction progress monitoring involves a set of inspection tasks with repetitive in-person observations on the site. The current manual inspection process in construction is time-consuming, inefficient and inconsistent mainly due to human limitations in the ability to persistently and accurately walkthrough the job site and observe the as-built status of which robots are considerably better. Enabling the process of visual inspection with a real-time and remote inspection capability using robots can provide more frequent and accessible construction progress data for inspectors to improve the quality of inspection and monitoring. Also, integrating remote inspection with an Augmented Reality (AR) platform can help the inspector to verify as-planned BIM data with the as-built status. This paper proposes a new approach to perform remote monitoring of the construction progress in real-time using a quadruped robot and an AR solution. The proposed computational framework in this study uses a cloud-based solution to integrate the quadruped robot’s control for remote navigation through the construction site with 360° live-stream video of the construction status, as well as a real-time AR solution to visualize and compare the as-built status with as-planned BIM geometry. The implementation of the proposed framework is discussed, and the developed framework is evaluated in two use cases through experimental investigations