Innovation in multifunctional bioabsorbable osteoconductive drug-releasing hard tissue fixation devices

We review in this paper the work performed by our group to develop multifunctional bioabsorbable ciprofloxacin releasing bone implants. Poly lactide-co-glycolide (PLGA 80/20 and polylactide (P(L/DL)LA 70/30) were used. Ciprofloxacin (CF) and bioactive glass (BaG) 13-93 were added. The mixture was then extruded and self-reinforced. CF release, mechanical strength, and the effect on S. epidermidis attachment and biofilm formation were evaluated. In rabbits, tissue reactions were assessed. Pull out strength was evaluated in cadaver bones. CF was released over 44 weeks (P(L/DL)LA) and 23–26 weeks (PLGA). Initial shear strength of the CF screws was 152 MPa (P(L/DL)LA) and 172 MPa (PLGA). Strength was retained for 12 weeks (P(L/DL)LA) and 9 weeks (PLGA). Histologically, CF releasing implants did not show much difference from control plain PLGA screws except for increased giant cells. CF miniscrews had lower pullout strength than the controls, but CF tacks had better values than controls. BaG led to a drop in pullout strength properties. Bacterial growth, attachment and biofilm formation on CF implants was significantly reduced when compared to controls. Accordingly, bioabsorbable multifunctional implants with appropriate CF release, mechanical, and biocompatibility properties are possible to develop and are considered appropriate to apply clinically.     

RELVEC — a tool for control system reliability analysis

This article will present a computerized reliability analysis tool for large control systems. It will also show a new dynamic representation of system structure. It enables us to model the physical system only once for any number of control tasks. The algorithm for computing minimal cut sets for the control tasks has been developed and automated. The result is RELVEC, an interactive computer program that performs reliability/availability calculation, sensitivity analysis and critical component identification. It can handle two repair policies and common mode failures. Reconfiquring of the physical system or the control tasks is simple. RELVEC is becoming an everyday tool in control system reliability analysis at VTT.     

Online Extrinsic Parameters Calibration for Stereovision Systems Used in Far-Range Detection Vehicle Applications

This paper presents a high-accuracy online calibration method for the absolute extrinsic parameters of a stereovision system that is suited for far-distance, vision-based vehicle applications. The method uses as prior knowledge the intrinsic parameters and the relative extrinsic parameters (relative position and orientation) of the two cameras, which are calibrated using offline procedures. These parameters remain unchanged if the two cameras are mounted on a rigid frame (stereo rig). The absolute extrinsic parameters define the position and orientation of the stereo system relative to a world coordinate system. They must be calibrated every time after mounting the stereo rig in the vehicle and are subject to changes due to static load factors for the used car setup. The proposed method is able to perform online the estimation of the absolute extrinsic parameters by driving the car on a flat and straight road, parallel with the longitudinal lane markers. The edge points of the longitudinal lane markers are extracted after a 2-D image classification process and reconstructed by stereovision in the stereo-rig coordinate system. After filtering out the noisy 3-D points, the normal vectors of the world coordinate system axes are estimated in the stereo-rig coordinate system by 3-D data fitting. The output of the method is the height and the orientation of the stereo cameras that are relative to the world coordinate system.     

Ubimedia based on readable and writable memory tags

Ubimedia is a concept where media files are embedded in everyday objects and the environment. We propose an approach where the user can read and write these files with his/her personal mobile phone simply by touching the physical objects. This facilitates easy access and storage of, e.g. video and audio files related to the physical object in question. This paper describes our work in developing a technical solution for ubimedia and studying user acceptance of forthcoming ubimedia services. Our technical development of the ubimedia concept has been focused on a mobile phone platform with a tag reader/writer, memory tags with large storage capacity, and the communication between the phone and the tags. Currently, the technical design is in test and evaluation phase. The preliminary results show that the concept works and it can be implemented technically. In parallel with the technical development, we have studied usage possibilities for ubimedia and user acceptance of future ubimedia services. User acceptance has been studied in a web survey and in user evaluations of proofs-of-concept. In addition, an ethical assessment has been carried out. The users appreciated especially the simplicity, speed, low cost and reliability of ubimedia. Ethical concerns were related to control over the download with regard to viruses and other unwanted content.     

Early alcoholic liver injury: formation of protein adducts with acetaldehyde and lipid peroxidation products, and expression of CYP2E1 and CYP3A

The formation of protein adducts with reactive aldehydes resulting from ethanol metabolism and lipid peroxidation has been suggested to play a role in the pathogenesis of alcoholic liver injury. To gain further insight on the contribution of such aldehydes in alcoholic liver disease, we have compared the appearance of acetaldehyde, malondialdehyde, and 4-hydroxynonenal adducts with the expression of cytochrome P-450IIE1, and cytochrome P-4503A enzymes in the liver of rats fed alcohol with a high-fat diet for 2 to 4 weeks according to the Tsukamoto-French procedure and in control rats (high-fat liquid diet or no treatment). Urine alcohol and serum aminotransferase levels were recorded, and the liver pathology was scored from 0 to 10 according to the presence of steatosis, inflammation, necrosis, and fibrosis. The ethanol treatment resulted in the accumulation of fat, mild necrosis and inflammation, and a mean liver pathology score of 3 (range: 1 to 5). Liver specimens from the ethanol-fed animals with early alcohol-induced liver injury were found to contain perivenular, hepatocellular acetaldehyde adducts. Malondialdehyde and 4-hydroxynonenal adducts were also present showing a more diffuse staining pattern with occasional sinusoidal reactions. In the control animals, a faint positive reaction for the hydroxynonenal adduct occurred in some of the animals fed the high fat diet, whereas no specific staining was observed in the livers from the animals receiving no treatment. Expression of both CYP2E1 and CYP3A correlated with the amount of protein adducts in the liver of alcohol-treated rats. Distinct CYP2E1-positive immunohistochemistry was seen in 3 of 7 of the ethanol-fed animals. In 5 of 7 of the ethanol-fed animals, the staining intensities for CYP3A markedly exceeded those obtained from the controls. The present findings indicate that acetaldehyde and lipid peroxidation-derived adducts are generated in the early phase of alcohol-induced liver disease. The formation of protein adducts appears to be accompanied by induction of both CYP2E1 and CYP3A.     

Physical and sensory properties of low-salt phosphate-free frankfurters composed with various ingredients

The physical properties and sensory attributes of phosphate-free frankfurters were examined using response surface methodology by varying the amounts of five compositional variables: salt, modified tapioca starch-, sodium citrate (NaC)- and wheat bran and fat in the batter. Altogether, 20 different types of frankfurters were prepared. When the frankfurters were made without phosphate, additional non-meat ingredients were needed at salt contents of less than 1.5%. Modified tapioca starch and sodium citrate decreased frying loss, with the former also improving water and fat binding.     

Analysis of the hypoxia-induced ADH2 promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast

We introduced a reporter gene system into Pichia stipitis using the gene for the artificial green fluorescent protein (GFP), variant yEGFP. This system was used to analyse hypoxia-dependent PsADH2 regulation. Reporter gene activity was only found under oxygen limitation on a fermentable carbon source. The promoter was not induced by oxygen limitation in the Crabtree-positive yeast Saccharomyces cerevisiae. Promoter deletions revealed that a region of 15 bp contained the essential site for hypoxic induction. This motif was different from the known hypoxia response elements of S. cerevisiae but showed some similarity to the mammalian HIF-1 binding site. Electrophoretic mobility shift assays demonstrated specific protein binding to this region under oxygen limitation. Similar to the S. cerevisiae heme sensor system, the promoter was induced by Co(2+). Cyanide was not able to mimic the effect of oxygen limitation. The activation mechanism of PsADH2 also, in this respect, has similarities to the mammalian HIF-1 system, which is inducible by Co(2+) but not by cyanide. Thus, the very first promoter analysis in P. stipitis revealed a hitherto unknown mechanism of oxygen sensing in yeast.     

Nanoparticles Interacting with Proteins and Cells: A Systematic Study of Protein Surface Charge Effects

Despite intense research on biological and biomedical applications of nanoparticles, our understanding of their basic interactions with the biological environment is still incomplete. Systematic variation of the physicochemical properties of the nanoparticles is widely seen as a promising strategy to obtain further insights. In view of the key role of the protein adsorption layer forming on nanoparticles in contact with biofluids, we systematically varied the surface charge of proteins adsorbing onto nanoparticles by chemical modification so as to examine the effect of Coulomb forces in modulating nano‐bio interactions. We chose human serum albumin (HSA) as a model protein and ultra‐small, negatively charged fluorescent gold nanoclusters (AuNCs) as model nanoparticles. By using fluorescence and CD spectroscopies, we measured binding affinities and structural changes upon binding of the HSA variants. The strengths of the protein‐nanoparticle interactions were found to change substantially upon modifying the surface charge of HSA. Furthermore, by using inductively coupled plasma optical emission spectroscopy, confocal fluorescence microscopy, scanning transmission electron microscopy and cell viability assays, we observed that cellular interactions of the AuNCs, including their adherence to cell membranes, uptake efficiency and cytotoxicity, depended markedly on the different surface charges of the HSA variants adsorbed onto the nanoparticles. These results illustrate vividly that the cellular responses to nanoparticle exposure depend on the specific properties of the proteins that adsorb onto nanoparticles from biofluids.     

Analysis of the RAP1 protein binding to homogeneous telomeric repeats in Saccharomyces castellii

The repressor activator protein 1 (RAP1) plays a role in telomere structure and function inS. cerevisiae. Here, the RAP1 homologue was identified and cloned from the budding yeast Saccharomyces castellii (scasRAP1). The scasRAP1 gene encodes a protein of 826 amino acids and shares an overall high degree of similarity with the S. cerevisiae RAP1 (scerRAP1). We demonstrate that the scasRAP1 is able to complement scerRAP1 in temperature-sensitive S. cerevisiae strains and is able to function as a regulator to maintain the original telomere lengths. Binding analyses of the E. coli-expressed scasRAP1 protein demonstrate that it needs two consecutive telomeric repeats in order to bind the S. castellii telomeric DNA sequences, and that it binds adjacent sites having a 16 bp centre-to-centre spacing. The binding affinity to telomeric DNA of several other yeasts is similar to that of scerRap1p. However, in contrast to scerRap1p, scasRap1p was found to bind the human telomeric sequence. Moreover, the scasRap1p was found to incorporate a variant repeat in its binding to the otherwise homogeneous telomeric DNA of S. castellii. This ability to bind various sites differing in DNA sequence indicates a high degree of adjustability in the binding of scasRap1p to DNA.     

Professionals’ knowledge and use of environmental assessment in an architectural competition

In early design phases, architects, landscape architects and urban planners are key actors whose decisions determine the environmental impact of planning and building projects. Environmental and sustainability assessment tools for buildings and neighbourhoods have been developed to promote sustainable building, but their usage has not been thoroughly evaluated. This study investigated self-reported knowledge and usage of such tools among competitors and jury group from 10 European countries involved in the international architectural competition ‘A New City Centre for Kiruna’ in Sweden. The questionnaire revealed that 13% used environmental assessment tools or management systems in the competition, although 47% had used them previously. Tool users reported greater knowledge of how to handle environmental impacts than non-users. However, the self-rated experience of handling various environmental impacts, in the competition and in general, was low for both groups. Nevertheless, the self-rated importance of environmental impacts was high among all participants. Based on this study, it is concluded that environmental assessment tools, issues and goals can be better integrated into the processes of early design in planning and building projects, and in architectural competitions. Furthermore, to limit environmental impacts in building and planning projects, professionals need to be educated about environmental strategies and solutions.