Chromatographic separation, fractionation and oxidation of selected beta-lactoglobulin peptides

The literature review elucidates the mechanism of oxidation in proteins and amino acids and gives an overview of the detection and analysis of protein oxidation products as well as information about ?-lactoglobulin and studies carried out on modifications of this protein under certain conditions. The experimental research included the fractionation of the tryptic peptides of ?-lactoglobulin using preparative-HPLC-MS and monitoring the oxidation process of these peptides via reverse phase-HPLC-UV. Peptides chosen to be oxidized were selected with respect to their amino acid content which were susceptible to oxidation and fractionated according to their m/z values. These peptides were: IPAVFK (m/z 674), ALPMHIR (m/z 838), LIVTQTMK (m/z 934) and VLVLDTDYK (m/z 1066). Even though it was not possible to solely isolate the target peptides due to co-elution of various fractions, the percentages of target peptides in the samples were satisfactory to carry out the oxidation procedure. IPAVFK and VLVLDTDYK fractions were found to yield the oxidation products reviewed in literature, however, unoxidized peptides were still present in high amounts after 21 days of oxidation. The UV data at 260 and 280 nm enabled to monitor both the main peptides and the oxidation products due to the absorbance of aromatic side-chains these peptides possess. ALPMHIR and LIVTQTMK fractions were oxidatively consumed rapidly and oxidation products of these peptides were observed even on day 0. High rates of depletion of these peptides were acredited to the presence of His (H) and sulfur-containing side-chains of Met (M). In conclusion, selected peptides hold the potential to be utilized as marker peptides in ?-lactoglobulin oxidation.

Building Information Modelling – Experts´ Views on Standardisation and Industry Deployment

The goal of the single building information model has existed for at least thirty years and various standards have been published leading up to the ten-year development of the Industry Foundation Classes. These have been initiatives from researchers, software developers and standards committees. Now large property owners are becoming aware of the benefits of moving IT tools from specific applications towards more comprehensive solutions. This study addresses the state of Building Information Models and the conditions necessary for them to become more widely used. It is a qualitative study based on information from a number of international experts and has asked a series of questions about the feasibility of BIMs, the conditions necessary for their success, and the role of standards with particular reference to the IFCs. Some key statements were distilled from the diverse answers received and indicate that BIM solutions appear too complex for many and may need to be applied in limited areas initially. Standards are generally supported but not applied rigorously and a range of these are relevant to BIM. Benefits will depend upon the building procurement methods used and there should be special roles within the project team to manage information. Case studies are starting to appear and these could be used for publicity. The IFCs are rather oversold and their complexities should be hidden within simple-to-use software. Inevitably major questions remain and property owners may be the key to answering some of these. A framework for presenting standards, backed up by case studies of successful projects, is the solution proposed to provide better information on where particular BIM standards and solutions should be applied in building projects.

Chemical approaches to penicillin allergy—IV. Binding of carrier receptor protein with penicillin and its analogues

The availability of electrophoretically homogeneous rabbit penicillin carrier receptor protein (CRP) by affinity chromatography afforded an idealin vitro system to calculate the thermodynamic parameters of binding of penicillin and analogues with CRP as well as competitive binding of such analogues with CRP in presence of14C-penicillin G. The kinetics of association of CRP with 7-deoxy penicillin which does not bind covalently with CRP have been studied through equilibrium dialysis with14C-7-deoxybenzyl penicillin and found to be K=2·79×106M−1.−ΔG=8·106 k cal/mole as well as fluorescence quenching studies with exciter λ 280 K=3·573×106M−1,−ΔG=8·239 k cal/mole. The fluorescence quenching studies have been extended to CRP-benzyl penicillin and CRP-6-aminopenicillanic acid (6APA) systems also. The fluorescence data with benzyl penicillin indicate two conformational changes in CRP—a fast change corresponding to the non-covalent binding to CRP with 7-deoxy penicillin and a slower change due to covalent bond formation. With 6-APA the first change is not observed but the conformational change corresponding to covalent binding is only seen. Competitive binding studies indicate that the order of binding of CRP with the analogues of penicillin is as follows: methicillin > 6APA > carbenicillin >o-nitrobenzyl penicillin > cloxacillin ≈ benzyl penicillin ≈ 6-phenyl acetamido penicillanyl alcohol ≈ 7 phenyl acetamido desacetoxy cephalosporanic acid ≈p-amino benzyl penicillin ≈p-nitro benzyl penicillin > ticarcillin >o-amino benzyl penicillin > amoxycillin > 7-deoxy benzyl penicillin > ampicillin.From these data it has been possible to delineate partially the topology of the penicillin binding cleft of the CRP as well as some of the functional groups in the cleft responsible for the binding process.

Compressible Boundary-Layer Swirling Flow in a Nozzle and a Diffuser With Highly Cooled Wall

The steady laminar compressible boundary-layer swirling flow with variable gas properties and mass transfer through a conical nozzle, and a diffuser with a highly cooled wall has been studied. The partial differential equations governing the nonsimilar flow have been transformed to a system of coordinates using modified Lees transformation. The resulting equations are transformed into coordinates having finite ranges by means of a transformation which maps an infinite region into a finite region. The ensuing equations are then solved numerically using an implicit finite-difference scheme. The results indicate that the variation of the density-viscosity product across the boundary layer and mass transfer have strong effect on the skin friction and heat transfer. Separationless flow along the entire length of the diffuser can be obtained by applying suction. The results are found to be in good agreement with those of the local nonsimilarity method but they differ appreciably from those of the local similarity method.

Dimensions of University Student Learning in Medicine and Pharmacy

Study orientations in higher education consist of various dimensions, such as approaches to learning, conceptions of learning and knowledge (i.e. epistemologies), self-regulation, and motivation. They have also been measured in different ways. The main orientations typically reported are reproducing and meaning orientations. The present study explored dimensions of study orientations, focusing in particular on pharmacy and medicine. New versions of self-report instruments were developed and tested in various contexts and in two countries. Furthermore, the linkages between study orientations and students epistemological development were explored. The context of problem-based (PBL) small groups was investigated in order to better understand how collaboration contributes to the quality of learning. The participants of Study I (n=66) were pharmacy students, who were followed during a three-year professionally oriented program in terms of their study orientations and epistemologies. A reproducing orientation to studying diminished during studying, whereas only a few students maintained their original level of meaning orientation. Dualism was found to be associated with a reproducing orientation. In Study II practices associated with deep and surface approaches to learning were measured in two differing ways, in order to better distinguish between what students believed to be useful in studying, and the extent to which they applied their beliefs to practice when preparing for examinations. Differences between domains were investigated by including a sample of Finnish and Swedish medical students (n=956) and a Finnish non-medical sample of university students (n=865). Memorizing and rote learning appeared as differing components of a surface approach to learning, while understanding, relating, and critical evaluation of knowledge emerged as aspects of a deep approach to learning. A structural model confirmed these results in both student samples. Study III explored a wide variety of dimensions of learning in medical education. Swedish medical students (n=280) answered the questionnaire. The deep approach to learning was strongly related to collaboration and reflective learning, whereas the surface approach was associated with novice-like views of knowledge and the valuing of certain and directly applicable knowledge. PBL students aimed at understanding, but also valued the role of memorization. Study IV investigated 12 PBL tutorial groups of students (n=116) studying microbiology and pharmacology in a medical school. The educational application was expected to support a deep approach to learning: Group members course grades in a final examination were related to the perceived functioning of the PBL tutorial groups. Further, the quality of cases that had been used as triggers for learning, was associated with the quality of small group functioning. New dimensions of study orientations were discovered. In particular, novel, finer distinctions were found within the deep approach component. In medicine, critical evaluation of knowledge appeared to be less valued than understanding and relating. Further, collaboration appeared to be closely related to the deep approach, and it was also important in terms of successful PBL studying. The results of the studies confirmed the previously found associations between approaches to learning and study success, but showed interesting context- and subgroup-related differences in this respect. Students ideas about the nature of knowledge and their approaches to learning were shown to be closely related. The present study expanded our understanding of the dimensions of study orientations, of their development, and their contextual variability in pharmacy and medicine.

Accelerometer-Based Force Balance for High Enthalpy Facilities

A single component accelerometer-based force balance is developed, calibrated, and used for high enthalpy applications. Functionality of this force balance, for such applications, is demonstrated for the first time during high enthalpy tests in a newly established free piston-driven shock tunnel, HST3, using a 60 degrees apex angle blunt cone model at 0 degrees angle of incidence. Usefulness of this force balance is also confirmed, for much complicated high enthalpy flow situations, during the drag reduction studies with counterflow supersonic jet from the stagnation point.

Temperature response of trehalase from a mesophilic (Neurospora crassa) and a thermophilic (Thermomyces lanuginosus) fungus

The purified trehalases of the mesophilic fungus, Neurospora crassa, and the thermophilic fungus, Thermomyces lanuginosus, had similar temperature and pH optima for activity, but differed in molecular weight, electrophoretic mobility and Michaelis constant. At lower concentration, trehalases from both fungi were inactivated to similar extent at 60°C. While purified trehalase of T. lanuginosus was afforded protection against heat-inactivation by proteinaceous protective factor(s) present in mycelial extracts, by bovine serum albumin and by casein, these did not afford protection to N. crassa trehalase against heat inactivation. Both trehalases exhibited discontinuous Arrhenius plots with temperature of discontinuity at 40°C. The activation energy calculated from the slope of the Arrhenius plot was higher for the T. lanuginosus enzyme. The plots of apparent K m versus 1/T for trehalases of N. crassa and T. lanuginosus were linear from 30° to 60°C. The results show that purified trehalases of the mesophilic and the thermophilic fungus are distinct. Although, these exhibit similar thermostability of their catalytic function at low concentration, distinctive thermal stability characteristics of thermophilic enzyme become apparent at high protein concentration. This could be brought about in the cell by the enzyme itself, or by other proteins.

Nucleophilic attacks of 1,2-diaminoethane on MeCN ligands: synthesis, x-ray structure, and spectral and electrochemical properties of [Ru(.mu.-O)(.mu.-O2CAr)2[NH2CH2CH2NHC(Me)NH]2(PPh3)2](ClO4)2(Ar = C6H4-p-X; X = H, Me, OMe, Cl)

The diruthenium(III) complex [Ru2O(O2CAr)2(MeCN)4(PPh3)2](ClO4)2 (1), on reaction with 1,2-diaminoethane (en) in MeOH at 25-degrees-C, undergoes nucleophilic attacks at the carbon of two facial MeCN ligands to form [(Ru2O)-O-III(O2CAr)2-{NH2CH2CH2NHC(Me)NH}2(PPh3)2](ClO4)2 (2) (Ar = C6H4-p-X, X = H, Me, OMe, Cl) containing two seven-membered amino-amidine chelating ligands. The molecular structure of 2 with Ar = C6H4-p-OMe was determined by X-ray crystallography. Crystal data are as follows: triclinic, P1BAR, a = 13.942 (5) angstrom, b = 14.528 (2) angstrom, c = 21.758 (6) angstrom, alpha = 109.50 (2)-degrees, beta = 92.52 (3)-degrees, gamma = 112.61 (2)-degrees, V = 3759 (2) angstrom 3, and Z = 2. The complex has an {Ru2(mu-O)(mu-O2CAr2)2(2+)} core. The Ru-Ru and average Ru-O(oxo) distances and the Ru-O-Ru angle are 3.280 (2) angstrom, 1.887 [8] angstrom, and 120.7 (4)-degrees, respectively. The amino group of the chelating ligand is trans to the mu-oxo ligand. The nucleophilic attacks take place on the MeCN ligands cis to the mu-oxo ligand. The visible spectra of 2 in CHCl3 display an absorption band at 565 nm. The H-1 NMR spectra of 2 in CDCl3 are indicative of the formation of an amino-amidine ligand. Complex 2 exhibits metal-centered quasireversible one-electron oxidation and reduction processes in the potential ranges +0.9 to +1.0 V and -0.3 to -0.5 V (vs SCE), respectively, involving the Ru(III)2/Ru(III)Ru(IV) and Ru(III)2/Ru(II)Ru(III) redox couples in CH2Cl2 containing 0.1 M TBAP. The mechanistic aspects of the nucleophilic reaction are discussed.

A mathematical analysis of nonlinear waves in a fluid filled visco-elastic tube

Our investigations in this paper are centred around the mathematical analysis of a ldquomodal waverdquo problem. We have considered the axisymmetric flow of an inviscid liquid in a thinwalled viscoelastic tube under certain simplifying assumptions. We have first derived the propagation space equations in the long wave limit and also given a general procedure to derive these equations for arbitrary wave length, when the flow is irrotational. We have used the method of operators of multiple scales to derive the nonlinear Schrödinger equation governing the modulation of periodic waves and we have elaborated on the ldquolong modulated wavesrdquo and the ldquomodulated long wavesrdquo. We have also examined the existence and stability of Stokes waves in this system. This is followed by a discussion of the progressive wave solutions of the long wave equations. One of the most important results of our paper is that the propagation space equations are no longer partial differential equations but they are in terms of pseudo-differential operators.Die vorliegenden Untersuchungen beziehen sich auf die mathematische Behandlung des ldquorModalwellenrdquo-Problems. Die achsensymmetrische Strömung einer nichtviskosen Flüssigkeit in einem dünnwandigen viskoelastischen Rohr, unter bestimmten vereinfachenden Annahmen, wird betrachtet. Zuerst werden die Gleichungen des Ausbreitungsraumes im Langwellenbereich abgeleitet und eine allgemeine Methode zur Herleitung dieser Gleichungen für beliebige Wellenlängen bei nichtrotierender Strömung angegeben. Eine Operatorenmethode mit multiplem Maßstab wird verwendet zur Herleitung der nichtlinearen Schrödinger-Gleichung für die Modulation der periodischen Wellen, und die ldquorlangmodulierten Wellenrdquo sowie die ldquormodulierten Langwellenrdquo werden aufgezeigt. Weiters wird die Existenz und die Stabilität der Stokes-Wellen im System untersucht. Anschließend werden die progressiven Wellenlösungen der Langwellengleichungen diskutiert. Eines der wichtigsten Ergebnisse dieser Arbeit ist, daß die Gleichungen des Ausbreitungsraumes keine partiellen Differentialgleichungen mehr sind, sondern Ausdrücke von Pseudo-Differentialoperatoren.

Rapid synthesis of room temperature ferromagnetic Ag-doped LaMnO3 perovskite phases by the solution combustion method

We report the rapid solution combustion synthesis and characterization of Ag-substituted LaMnO3 phases at relatively low temperature using oxalyl dihydrazide, as fuel. Structural parameters were refined by the Rietveld method using powder X-ray diffraction data. While the parent LaMnO3 crystallizes in the orthorhombic structure, the Ag-substituted compounds crystallize in the rhombohedral symmetry. On increasing Ag-content, unit cell volume and Mn-O-Mn bond angle decreases. The Fourier transform infra red spectrum shows two absorption bands corresponding to Mn-O stretching vibration (v(s) mode) and Mn-O-Mn deformation vibration (v(b) mode) around 600 cm(-1) and 400 cm(-1) for the compositions x = 0.0, 0.05 and 0.10, respectively. Electrical resistivity measurements reveal that composition-controlled metal to insulator transition, with the maximum metal to insulator being 280 K for the composition La0.75Ag0.25MnO3. Increase in magnetic moment was observed with increase in Ag-content. The maximum magnetic moment of 35 emu/g was observed for the composition La0.80Ag0.20MnO3. (C) 2010 Elsevier Ltd. All rights reserved.

Pain measurement and management in elderly patients : Clinical studies in long term hospital care and after cardiac surgery

The proportion of patients over 75 years of age, receiving all different types of healthcare, is constantly increasing. The elderly undergo surgery and anaesthetic procedures more often than middle-aged patients. Poor pain management in the elderly is still an issue. Although the elderly consumes the greatest proportion of prescribed medicines in Western Europe, most clinical pharmacological studies have been performed in healthy volunteers or middle-aged patients. The aim of this study was to investigate pain measurement and management in cognitively impaired patients in long term hospital care and in cognitively normal elderly patients after cardiac surgery. This thesis incorporated 366 patients, including 86 home-dwelling or hospitalized elderly with chronic pain and 280 patients undergoing cardiac surgery with acute pain. The mean age of patients was 77 (SD ± 8) years and approximately 8400 pain measurements were performed with four pain scales: Verbal Rating Scale (VRS), the Visual Analogue Scale (VAS), the Red Wedge Scale (RWS), and the Facial Pain Scale (FPS). Cognitive function, depression, functional ability in daily life, postoperative sedation and postoperative confusion were assessed with MMSE, GDS, Barthel Index, RASS, and CAM-ICU, respectively. The effects and plasma concentrations of fentanyl and oxycodone were measured in elderly (≥ 75 years) and middle-aged patients (≤ 60 years) and the opioid-sparing effect of pregabalin was studied after cardiac surgery. The VRS pain scores after movement correlated with the Barthel Index. The VRS was most successful in the groups of demented patients (MMSE 17-23, 11-16 and ≤ 10) and in elderly patients on the first day after cardiac surgery. The elderly had a higher plasma concentration of fentanyl at the end of surgery than younger patients. The plasma concentrations of oxycodone were comparable between the groups. Pain intensity on the VRS was lower and the sedation scores were higher in the elderly. Total oxycodone consumption during five postoperative days was reduced by 48% and the CAM-ICU scores were higher on the first postoperative day in the pregabalin group. The incidence of postoperative pain during movement was lower in the pregabalin group three months after surgery. This investigation demonstrates that chronic pain did not seem to impair daily activities in home-dwelling Finnish elderly. The VRS appeared to be applicable for elderly patients with clear cognitive dysfunction (MMSE ≤17) and it was the most feasible pain scale for the early postoperative period after cardiac surgery. After cardiac surgery, plasma concentrations of fentanyl in elderly were elevated, although oxycodone concentrations were at similar level compared to middle-aged patients. The elderly had less pain and were more sedated after doses of oxycodone. Therefore, particular attention must be given to individual dosing of the opioids in elderly surgical patients, who often need a smaller amount for adequate analgesia than middle-aged patients. The administration of pregabalin reduced postoperative oxycodone consumption after cardiac surgery. Pregabalin-treated patients had less confusion, and additionally to less postoperative pain on the first postoperative day and during movement at three months post-surgery. Pregabalin might be a new alternative as analgesic for acute postoperative and chronic pain management in the elderly. Its clinical role and safety remains to be verified in large-scale randomized and controlled studies. In the future, many clinical trials in the older category of patients will be needed to facilitate improvements in health care methods.

New pathway for the biodegradation of indole in Aspergillus niger

Indole and its derivatives form a class of toxic recalcitrant environmental pollutants. The growth of Aspergillus niger was inhibited by very low concentrations (0.005 to 0.02%) of indole, even when 125- to 500-fold excess glucose was present in the medium. When 0.02% indole was added, the fungus showed a lag phase for about 30 h and the uptake of glucose was inhibited. Indole was metabolized by a new pathway via indoxyl (3-hydroxyindole), N-formylanthranilic acid, anthranilic acid,2,3-dihydroxybenzoic acid, and catechol, which was further degraded by ortho cleavage. The enzymes N-formylanthranilate deformylase, anthranilate hydroxylase, 2,3-dihydroxybenzoate decarboxylase, and catechol dioxygenase were induced by indole as early as after 5 h of growth, and their activities were demonstrated in a cell-free system.

Naudanlihan jalostusketjun hiilijalanjälki Suomessa

Kirjallisuustutkimuksen tavoitteena oli perehtyä kasvihuoneilmiön taustoihin ja kartoittaa aiempia tutkimuksia naudan- ja muiden lihatuotteiden kasvihuonekaasupäästöistä. Lisäksi kirjallisuustutkimuksessa perehdyttiin aiemmissa tutkimuksissa elintarvikkeiden hiilijalanjäljen laskemisessa sovellettuun elinkaarianalyysiin ISO 14040-standardin mukaisesti. Kokeellisen osion tavoitteena oli määrittää naudanlihan hiilijalanjälki Suomessa maatilan portilta kuluttajan ruokapöytään. Tavoitteena oli myös ymmärtää jalostusketjun päästöjen merkitys verrattuna koko naudanlihan tuotantoketjuun ja määrittää jalostusketjun vaiheiden merkitys ketjussa. Työn toiminnallisena yksikkönä toimi kilo naudanlihaa. Työ toteutettiin perehtymällä yksityiskohtaisesti yhteen naudanlihan jalostusketjuun Suomessa. Päästöt laskettiin todellisten yhteistyöyritykseltä saatujen prosessitietojen perusteella. Tiedot kerättiin tiedonkeruulomakkeella vierailemalla yhteistyöyrityksen kahdessa tuotantolaitoksessa ja täydentämällä tietoja haastatteluilla. Naudanlihan jalostusketjun päästöt olivat 1240 g CO2-ekv/lihakilo. Eniten päästöjä tuottivat jalostusvaihe (310 g CO2-ekv/lihakilo), teurastus (280 g CO2-ekv/lihakilo) ja lihatuotteiden kuljetus kuluttajalle (210 g CO2-ekv/lihakilo). Koko naudanlihan tuotantoketjusta jalostusketjun päästöt muodostivat alle 4 %, sillä syntymästä maatilan portille syntyviksi päästöiksi laskettiin kirjallisuuden perusteella yli 30 000 g CO2-ekv/lihakilo. Jatkossa naudanlihan hiilijalanjälkeä voitaisiin pääasiassa pienentää kehittämällä prosessia maatilan portille asti. Tämän työn tulokset olivat hyvin samansuuruiset verrattuna aiempaan tutkimukseen broilerin jalostusketjun päästöistä Suomessa (Katajajuuri ym. 2008). Tämä vastasi ennakko-odotuksia, sillä jalostusketjujen vaiheissa ei ollut merkittäviä eroja. Aiempia tutkimuksia naudanlihan jalostusketjun päästöistä ei ollut saatavilla.