Effect of chemical conjugation of L-leucine to chitosan on the dispersibility and drug release of a dry powder inhaler nanoparticle formulation

Purpose: This study investigated the effect of chemical conjugation of the amino acid L-leucine to the polysaccharide chitosan on the dispersibility and drug release pattern of a polymeric nanoparticle (NP)-based controlled release dry powder inhaler (DPI) formulation. Methods: A chemical conjugate of L-leucine with chitosan was synthesized and characterized by Infrared (IR) Spectroscopy, Nuclear Magnetic Resonance (NMR) Spectroscopy, Elemental Analysis and X-ray Photoelectron Spectroscopy (XPS). Nanoparticles of both chitosan and its conjugate were prepared by a water-in-oil emulsification – glutaraldehyde cross-linking method using the antihypertensive agent, diltiazem (Dz) hydrochloride as the model drug. The surface morphology and particle size distribution of the nanoparticles were determined by Scanning Electron Microscopy (SEM) and Dynamic Light Scattering (DLS). The dispersibility of the nanoparticle formulation was analysed by a Twin Stage Impinger (TSI) with a Rotahaler as the DPI device. Deposition of the particles in the different stages was determined by gravimetry and the amount of drug released was analysed by UV spectrophotometry. The release profile of the drug was studied in phosphate buffered saline at 37 ⁰C and analyzed by UV spectrophotometry. Results: The TSI study revealed that the fine particle fractions (FPF), as determined gravimetrically, for empty and drug-loaded conjugate nanoparticles were significantly higher than for the corresponding chitosan nanoparticles (24±1.2% and 21±0.7% vs 19±1.2% and 15±1.5% respectively; n=3, p<0.05). The FPF of drug-loaded chitosan and conjugate nanoparticles, in terms of the amount of drug determined spectrophotometrically, had similar values (21±0.7% vs 16±1.6%). After an initial burst, both chitosan and conjugate nanoparticles showed controlled release that lasted about 8 to 10 days, but conjugate nanoparticles showed twice as much total drug release compared to chitosan nanoparticles (~50% vs ~25%). Conjugate nanoparticles also showed significantly higher dug loading and entrapment efficiency than chitosan nanoparticles (conjugate: 20±1% & 46±1%, chitosan: 16±1% & 38±1%, n=3, p<0.05). Conclusion: Although L-leucine conjugation to chitosan increased dispersibility of formulated nanoparticles, the FPF values are still far from optimum. The particles showed a high level of initial burst release (chitosan, 16% and conjugate, 31%) that also will need further optimization.

Exploring schema-driven differences in situation awareness between road users : an on-road study of driver, cyclist and motorcyclist situation awareness

Collisions between different road users make a substantial contribution to road trauma. Although evidence suggests that different road users interpret the same road situations differently, it is not clear how road users' situation awareness differs, nor is it clear which differences might lead to conflicts. This article presents the findings from an on-road study conducted to examine driver, motorcyclist and cyclist situation awareness in different road environments. The findings suggest that, in addition to minor differences in the structure of different road users' situation awareness (i.e. amount of information and how it is integrated), the actual content of situation awareness in terms of road user schemata, the resulting interaction with the world and the information underpinning situation awareness is markedly different. Further examination indicates that the differences are likely to be compatible along arterial roads, shopping strips and at roundabouts, but that they may create conflicts between different road users at intersections. Interventions designed to support compatible situation awareness and behaviour between different road users are discussed. Practitioner Summary: Incompatible situation awareness plays a key role in collisions between different road users (e.g. drivers and motorcyclists). This on-road study examined situation awareness in drivers, motorcyclists and cyclists, identifying the key differences and potential conflicts that arise. The findings are used to propose interventions designed to enhance the compatibility of situation awareness between road users.

Enhanced geoproof : improved geographic assurance for data in the cloud

The geographic location of cloud data storage centres is an important issue for many organisations and individuals due to various regulations that require data and operations to reside in specific geographic locations. Thus, cloud users may want to be sure that their stored data have not been relocated into unknown geographic regions that may compromise the security of their stored data. Albeshri et al. (2012) combined proof of storage (POS) protocols with distance-bounding protocols to address this problem. However, their scheme involves unnecessary delay when utilising typical POS schemes due to computational overhead at the server side. The aim of this paper is to improve the basic GeoProof protocol by reducing the computation overhead at the server side. We show how this can maintain the same level of security while achieving more accurate geographic assurance.

Understanding the abnormal brain activity in epilepsy as a potential predictor of the onset of an epileptic seizure

The Brain Research Institute (BRI) uses various types of indirect measurements, including EEG and fMRI, to understand and assess brain activity and function. As well as the recovery of generic information about brain function, research also focuses on the utilisation of such data and understanding to study the initiation, dynamics, spread and suppression of epileptic seizures. To assist with the future focussing of this aspect of their research, the BRI asked the MISG 2010 participants to examine how the available EEG and fMRI data and current knowledge about epilepsy should be analysed and interpreted to yield an enhanced understanding about brain activity occurring before, at commencement of, during, and after a seizure. Though the deliberations of the study group were wide ranging in terms of the related matters considered and discussed, considerable progress was made with the following three aspects. (1) The science behind brain activity investigations depends crucially on the quality of the analysis and interpretation of, as well as the recovery of information from, EEG and fMRI measurements. A number of specific methodologies were discussed and formalised, including independent component analysis, principal component analysis, profile monitoring and change point analysis (hidden Markov modelling, time series analysis, discontinuity identification). (2) Even though EEG measurements accurately and very sensitively record the onset of an epileptic event or seizure, they are, from the perspective of understanding the internal initiation and localisation, of limited utility. They only record neuronal activity in the cortical (surface layer) neurons of the brain, which is a direct reflection of the type of electrical activity they have been designed to record. Because fMRI records, through the monitoring of blood flow activity, the location of localised brain activity within the brain, the possibility of combining fMRI measurements with EEG, as a joint inversion activity, was discussed and examined in detail. (3) A major goal for the BRI is to improve understanding about ``when'' (at what time) an epileptic seizure actually commenced before it is identified on an eeg recording, ``where'' the source of this initiation is located in the brain, and ``what'' is the initiator. Because of the general agreement in the literature that, in one way or another, epileptic events and seizures represent abnormal synchronisations of localised and/or global brain activity the modelling of synchronisations was examined in some detail. References C. M. Michel, G. Thut, S. Morand, A. Khateb, A. J. Pegna, R. Grave de Peralta, S. Gonzalez, M. Seeck and T. Landis, Electric source imaging of human brain functions, Brain Res. Rev. , 36 (2--3), 2001, 108--118. doi:10.1016/S0165-0173(01)00086-8 S. Ogawa, R. S. Menon, S. G. Kim and K. Ugurbil, On the characteristics of functional magnetic resonance imaging of the brain, Annu. Rev. Bioph. Biom. , 27 , 1998, 447--474. doi:10.1146/annurev.biophys.27.1.447 C. D. Binnie and H. Stefan, Modern electroencephalography: its role in epilepsy management, Clin. Neurophysiol. , 110 (10), 1999, 1671--1697. doi:10.1016/S1388-2457(99)00125-X J. X. Tao, A. Ray, S. Hawes-Ebersole and J. S. Ebersole, Intracranial eeg substrates of scalp eeg interictal spikes, Epilepsia , 46 (5), 2005, 669--76. doi:10.1111/j.1528-1167.2005.11404.x S. Ogawa, D. W. Tank, R. Menon, J. M. Ellermann, S. G. Kim, H. Merkle and K. Ugurbil, Intrinsic signal changes accompanying sensory stimulation: Functional brain mapping with magnetic resonance imaging, P. Natl. Acad. Sci. USA , 89 (13), 1992, 5951--5955. doi:10.1073/pnas.89.13.5951 J. Engel Jr., Report of the ilae classification core group, Epilepsia , 47 (9), 2006, 1558--1568. doi:10.1111/j.1528-1167.2006.00215.x L. Lemieux, A. Salek-Haddadi, O. Josephs, P. Allen, N. Toms, C. Scott, K. Krakow, R. Turner and D. R. Fish, Event-related fmri with simultaneous and continuous eeg: description of the method and initial case r port, NeuroImage , 14 (3), 2001, 780--7. doi:10.1006/nimg.2001.0853 P. Federico, D. F. Abbott, R. S. Briellmann, A. S. Harvey and G. D. Jackson, Functional mri of the pre-ictal state, Brain , 128 (8), 2005, 1811-7. doi:10.1093/brain/awh533 C. S. Hawco, A. P. Bagshaw, Y. Lu, F. Dubeau and J. Gotman, bold changes occur prior to epileptic spikes seen on scalp eeg, NeuroImage , 35 (4), 2007, 1450--1458. doi:10.1016/j.neuroimage.2006.12.042 F. Moeller, H. R. Siebner, S. Wolff, H. Muhle, R. Boor, O. Granert, O. Jansen, U. Stephani and M. Siniatchkin, Changes in activity of striato-thalamo-cortical network precede generalized spike wave discharges, NeuroImage , 39 (4), 2008, 1839--1849. doi:10.1016/j.neuroimage.2007.10.058 V. Osharina, E. Ponchel, A. Aarabi, R. Grebe and F. Wallois, Local haemodynamic changes preceding interictal spikes: A simultaneous electrocorticography (ecog) and near-infrared spectroscopy (nirs) analysis in rats, NeuroImage , 50 (2), 2010, 600--607. doi:10.1016/j.neuroimage.2010.01.009 R. S. Fisher, W. Boas, W. Blume, C. Elger, P. Genton, P. Lee and J. Engel, Epileptic seizures and epilepsy: Definitions proposed by the international league against epilepsy (ilae) and the international bureau for epilepsy (ibe), Epilepsia , 46 (4), 2005, 470--472. doi:10.1111/j.0013-9580.2005.66104.x H. Berger, Electroencephalogram in humans, Arch. Psychiat. Nerven. , 87 , 1929, 527--570. C. M. Michel, M. M. Murray, G. Lantz, S. Gonzalez, L. Spinelli and R. G. de Peralta, eeg source imaging, Clin. Neurophysiol. , 115 (10), 2004, 2195--2222. doi:10.1016/j.clinph.2004.06.001 P. L. Nunez and R. B. Silberstein, On the relationship of synaptic activity to macroscopic measurements: Does co-registration of eeg with fmri make sense?, Brain Topogr. , 13 (2), 2000, 79--96. doi:10.1023/A:1026683200895 S. Ogawa, T. M. Lee, A. R. Kay and D. W. Tank, Brain magnetic resonance imaging with contrast dependent on blood oxygenation, P. Natl. Acad. Sci. USA , 87 (24), 1990, 9868--9872. doi:10.1073/pnas.87.24.9868 J. S. Gati, R. S. Menon, K. Ugurbil and B. K. Rutt, Experimental determination of the bold field strength dependence in vessels and tissue, Magn. Reson. Med. , 38 (2), 1997, 296--302. doi:10.1002/mrm.1910380220 P. A. Bandettini, E. C. Wong, R. S. Hinks, R. S. Tikofsky and J. S. Hyde, Time course EPI of human brain function during task activation, Magn. Reson. Med. , 25 (2), 1992, 390--397. K. K. Kwong, J. W. Belliveau, D. A. Chesler, I. E. Goldberg, R. M. Weisskoff, B. P. Poncelet, D. N. Kennedy, B. E. Hoppelm, M. S. Cohen and R. Turner, Dynamic magnetic resonance imaging of human brain activity during primary sensory stimulation, P. Natl. Acad. Sci. USA , 89 (12), 1992, 5675--5679. doi:10.1073/pnas.89.12.5675 J. Frahm, K. D. Merboldt and W. Hnicke, Functional mri of human brain activation at high spatial resolution, Magn. Reson. Med. , 29 (1), 1993, 139--144. P. A. Bandettini, A. Jesmanowicz, E. C. Wong and J. S. Hyde, Processing strategies for time-course data sets in functional MRI of the human brain, Magn. Reson. Med. , 30 (2), 1993, 161--173. K. J. Friston, P. Jezzard and R. Turner, Analysis of functional MRI time-series, Hum. Brain Mapp. , 1 (2), 1994, 153--171. B. Biswal, F. Z. Yetkin, V. M. Haughton and J. S. Hyde, Functional connectivity in the motor cortex of resting human brain using echo-planar mri, Mag. Reson. Med. , 34 (4), 1995, 537--541. doi:10.1002/mrm.1910340409 K. J. Friston, J. Ashburner, C. D. Frith, J. Poline, J. D. Heather and R. S. J. Frackowiak, Spatial registration and normalization of images, Hum. Brain Mapp. , 3 (3), 1995, 165--189. K. J. Friston, S. Williams, R. Howard, R. S. Frackowiak and R. Turner, Movement-related effects in fmri time-series, Magn. Reson. Med. , 35 (3), 1996, 346--355. G. H. Glover, T. Q. Li and D. Ress, Image-based method for retrospective correction of physiological motion effects in fmri: Retroicor, Magn. Reson. Med. , 44 (1), 2000, 162--167. doi:10.1002/1522-2594(200007)44:13.0.CO;2-E K. J. Friston, O. Josephs, G. Rees and R. Turner, Nonlinear event-related responses in fmri, Magn. Reson. Med. , 39 (1), 1998, 41--52. doi:10.1002/mrm.1910390109 K. Ugurbil, L. Toth and D. Kim, How accurate is magnetic resonance imaging of brain function?, Trends Neurosci. , 26 (2), 2003, 108--114. doi:10.1016/S0166-2236(02)00039-5 D. S. Kim, I. Ronen, C. Olman, S. G. Kim, K. Ugurbil and L. J. Toth, Spatial relationship between neuronal activity and bold functional mri, NeuroImage , 21 (3), 2004, 876--885. doi:10.1016/j.neuroimage.2003.10.018 A. Connelly, G. D. Jackson, R. S. Frackowiak, J. W. Belliveau, F. Vargha-Khadem and D. G. Gadian, Functional mapping of activated human primary cortex with a clinical mr imaging system, Radiology , 188 (1), 1993, 125--130. L. Allison, Hidden Markov Models, Technical Report , School of Computer and Software Engineering, Monash University, 2000. R. J. Elliott, L. Aggoun and J.B. Moore, Hidden Markov Models: Estimation and Control, Appl. Math.-Czech. , 2004. B. Bhavnagri, Discontinuities of plane functions projected from a surface with methods for finding these , Technical Report, 2009. B. Bhavnagri, Computer Vision using Shape Spaces , Technical Report,1996, University of Adelaide. B. Bhavnagri, A method for representing shape based on an equivalence relation on polygons, Pattern Recogn. , 27 (2), 1994, 247--260. doi:10.1016/0031-3203(94)90057-4 D. F. Abbott, A. B. Waites, A. S. Harvey and G. D. Jackson, Exploring epileptic seizure onset with fmri, NeuroImage , 36(S1) (344TH-PM), 2007. M. C. Mackey and L. Glass, Oscillation and chaos in physiological control systems, Science , 197 , 1977, 287--289. S. H. Strogatz, SYNC - The Emerging Science of Spontaneous Order , Theia, New York, 2003. J. W. Kim, J. A. Roberts and P. A. Robinson, Dynamics of epileptic seizures: Evolution, spreading, and suppression, J. Theor. Biol. , 257 (4), 2009, 527--532. doi:10.1016/j.jtbi.2008.12.009 Y. Kuramoto, T. Aoyagi, I. Nishikawa, T. Chawanya T and K. Okuda, Neural network model carrying phase information with application to collective dynamics, J. Theor. Phys. , 87 (5), 1992, 1119--1126. V. B. Mountcastle, The columnar organization of the neocortex, Brain , 120 (4), 1997, 701. doi:10.1093/brain/120.4.701 F. L. Silva, W. Blanes, S. N. Kalitzin, J. Parra, P. Suffczynski and D. N. Velis, Epilepsies as dynamical diseases of brain systems: Basic models of the transition between normal and epileptic activity, Epilepsia , 44 (12), 2003, 72--83. F. H. Lopes da Silva, W. Blanes, S. N. Kalitzin, J. Parra, P. Suffczynski and D. N. Velis, Dynamical diseases of brain systems: different routes to epileptic seizures, ieee T. Bio-Med. Eng. , 50 (5), 2003, 540. L.D. Iasemidis, Epileptic seizure prediction and control, ieee T. Bio-Med. Eng. , 50 (5), 2003, 549--558. L. D. Iasemidis, D. S. Shiau, W. Chaovalitwongse, J. C. Sackellares, P. M. Pardalos, J. C. Principe, P. R. Carney, A. Prasad, B. Veeramani, and K. Tsakalis, Adaptive epileptic seizure prediction system, ieee T. Bio-Med. Eng. , 50 (5), 2003, 616--627. K. Lehnertz, F. Mormann, T. Kreuz, R.G. Andrzejak, C. Rieke, P. David and C. E. Elger, Seizure prediction by nonlinear eeg analysis, ieee Eng. Med. Biol. , 22 (1), 2003, 57--63. doi:10.1109/MEMB.2003.1191451 K. Lehnertz, R. G. Andrzejak, J. Arnhold, T. Kreuz, F. Mormann, C. Rieke, G. Widman and C. E. Elger, Nonlinear eeg analysis in epilepsy: Its possible use for interictal focus localization, seizure anticipation, and prevention, J. Clin. Neurophysiol. , 18 (3), 2001, 209. B. Litt and K. Lehnertz, Seizure prediction and the preseizure period, Curr. Opin. Neurol. , 15 (2), 2002, 173. doi:10.1097/00019052-200204000-00008 B. Litt and J. Echauz, Prediction of epileptic seizures, Lancet Neurol. , 1 (1), 2002, 22--30. doi:10.1016/S1474-4422(02)00003-0 M. M{a}kiranta, J. Ruohonen, K Suominen, J. Niinim{a}ki, E. Sonkaj{a}rvi, V. Kiviniemi, T. Sepp{a}nen, S. Alahuhta, V. J{a}ntti and O. Tervonen, {bold} signal increase preceeds eeg spike activity--a dynamic penicillin induced focal epilepsy in deep anesthesia, NeuroImage , 27 (4), 2005, 715--724. doi:10.1016/j.neuroimage.2005.05.025 K. Lehnertz, F. Mormann, H. Osterhage, A. M{u}ller, J. Prusseit, A. Chernihovskyi, M. Staniek, D. Krug, S. Bialonski and C. E. Elger, State-of-the-art of seizure prediction, J. Clin. Neurophysiol. , 24 (2), 2007, 147. doi:10.1097/WNP.0b013e3180336f16 F. Mormann, T. Kreuz, C. Rieke, R. G. Andrzejak, A. Kraskov, P. David, C. E. Elger and K. Lehnertz, On the predictability of epileptic seizures, Clin. Neurophysiol. , 116 (3), 2005, 569--587. doi:10.1016/j.clinph.2004.08.025 F. Mormann, R. G. Andrzejak, C. E. Elger and K. Lehnertz, Seizure prediction: the long and winding road, Brain , 130 (2), 2007, 314--333. doi:10.1093/brain/awl241 Z. Rogowski, I. Gath and E. Bental, On the prediction of epileptic seizures, Biol. Cybern. , 42 (1), 1981, 9--15. Y. Salant, I. Gath, O. Henriksen, Prediction of epileptic seizures from two-channel eeg, Med. Biol. Eng. Comput. , 36 (5), 1998, 549--556. doi:10.1007/BF02524422 J. Gotman and D.J. Koffler, Interictal spiking increases after seizures but does not after decrease in medication, Evoked Potential , 72 (1), 1989, 7--15. J. Gotman and M. G. Marciani, Electroencephalographic spiking activity, drug levels, and seizure occurence in epileptic patients, Ann. Neurol. , 17 (6), 1985, 59--603. A. Katz, D. A. Marks, G. McCarthy and S. S. Spencer, Does interictal spiking change prior to seizures?, Electroen. Clin. Neuro. , 79 (2), 1991, 153--156. A. Granada, R. M. Hennig, B. Ronacher, A. Kramer and H. Herzel, Phase Response Curves: Elucidating the dynamics of couples oscillators, Method Enzymol. , 454 (A), 2009, 1--27. doi:10.1016/S0076-6879(08)03801-9 doi:10.1016/S0076-6879(08)03801-9 H. Kantz and T. Schreiber, Nonlinear time series analysis , 2004, Cambridge Univ Press. M. V. L. Bennett and R. S Zukin, Electrical coupling and neuronal synchronization in the mammalian brain, Neuron , 41 (4), 2004, 495 --511. doi:10.1016/S0896-6273(04)00043-1 L.D. Iasemidis, J. Chris Sackellares, H. P. Zaveri and W. J. Williams, Phase space topography and the Lyapunov exponent of electrocorticograms in partial seizures, Brain Topogr. , 2 (3), 1990, 187--201. doi:10.1007/BF01140588 M. Le Van Quyen, J. Martinerie, V. Navarro, M. Baulac and F. J. Varela, Characterizing neurodynamic changes before seizures, J. Clin. Neurophysiol. , 18 (3), 2001, 191. J. Martinerie, C. Adam, M. Le Van Quyen, M. Baulac, S. Clemenceau, B. Renault and F. J. Varela, Epileptic seizures can be anticipated by non-linear analysis, Nat. Med. , 4 (10), 1998, 1173--1176. doi:10.1038/2667 A. Pikovsky, M. Rosenblum, J. Kurths and R. C. Hilborn, Synchronization: A universal concept in nonlinear science, Amer. J. Phys. , 70 , 2002, 655. H. R. Wilson and J. D. Cowan, Excitatory and inhibitory interactions in localized populations of model neurons, Biophys. J. , 12 (1), 1972, 1--24. D. Cumin and C. P. Unsworth, Generalising the Kuramoto model for the study of neuronal synchronisation in the brain, Physica D , 226 (2), 2007, 181--196. doi:10.1016/j.physd.2006.12.004 F. K. Skinner, H. Bazzazi and S. A. Campbell, Two-cell to N-cell heterogeneous, inhibitory networks: Precise linking of multistable and coherent properties, J. Comput. Neurosci. , 18 (3), 2005, 343--352. doi:10.1007/s10827-005-0331-1 W. W. Lytton, Computer modelling of epilepsy, Nat. Rev. Neurosci. , 9 (8), 2008, 626--637. doi:10.1038/nrn2416 R. D. Traub, A. Bibbig, F. E. N. LeBeau, E. H. Buhl and M. A. Whittington, Cellular mechanisms of neuronal population oscillations in the hippocampus in vitro, Ann. Rev. , 2004. R. D. Traub, A. Draguhn, M. A. Whittington, T. Baldeweg, A. Bibbig, E. H. Buhl and D. Schmitz, Axonal gap junc ions between principal neurons: A novel source of network oscillations, and perhaps epileptogenesis., Rev. Neuroscience , 13 (1), 2002, 1. doi:10.1146/annurev.neuro.27.070203.144303 M. Scheffer, J. Bascompte, W. A. Brock, V. Brovkin, S. R. Carpenter, V. Dakos, H. Held, E. H. van Nes, M. Rietkerk and G. Sugihara, Early-warning signals for critical transitions, Nature , 461 (7260), 2009, 53--59. doi:10.1038/nature08227 K. Murphy, A Brief Introduction to Graphical Models and Bayesian Networks , 2008, http://www.cs.ubc.ca/murphyk/Bayes/bnintro.html . R. C. Bradley, An elementary

Molecular structural studies of the amorphous mineral pitticite Fe, AsO4, SO4, H2O

Some minerals are colloidal and show no X-ray diffraction patterns. Vibrational spectroscopy offers one of the few methods for the assessment of the structure of these types of mineral. Among this group of minerals is pitticite simply described as Fe, AsO4, SO4, H2O. The objective of this research is to determine the molecular structure of the mineral pitticite using vibrational spectroscopy. Raman microscopy offers a useful method for the analysis of such colloidal minerals. Raman and infrared bands are attributed to the , and water stretching vibrations. The Raman spectrum is dominated by a very intense sharp band at 983 cm−1 assigned to the symmetric stretching mode. A strong Raman band at 1041 cm−1 is observed and is assigned to the antisymmetric stretching mode. Low intensity Raman bands at 757 and 808 cm−1 may be assigned to the antisymmetric and symmetric stretching modes. Raman bands observed at 432 and 465 cm−1 are attributable to the doubly degenerate ν2(SO4)2- bending mode.

Stress effects caused by the expression of a mutant cellobiohydrolase I and proteasome inhibition in Trichoderma reesei Rut-C30

Trichoderma reesei Rut-C30 is used widely as an expression host for various gene products. We have explored cellular effects caused by the expression of a mutant form of cellobiohydrolase I (CBHI), the major secreted protein of T. reesei using biochemical and transcriptomic analyses and confocal laser scanning microscopy. The mutated CBHI was tagged fluorescently with Venus to establish the subcellular location of the fusion protein and its potential association with the proteasome, an organelle assigned for the disposal of misfolded proteins. Expression of the mutant CBHI in the high protein-secreting host Rut-C30 caused physiological changes in the fungal hyphae, affected protein secretion and elicited ER stress. A massive upregulation of UPR- and ERAD-related genes sec61, der1, uba1, bip1, pdi1, prp1, cxl1 and lhs1 was observed by qRT-PCR in the CBHIΔ4-Venus strain with four mutations introduced in the DNA encoding the core domain of CBHI. Further stress was applied to this strain by inhibiting function of the proteasome with MG132 (N-benzoylcarbonyl(Cbz)-Leu-Leu-leucinal). The effect of MG132 was found to be specific to the proteasome-associated genes. There are no earlier reports on the effect of proteasome inhibition on protein quality control in filamentous fungi. Confocal fluorescence microscopy studies suggested that the mutant CBHI accumulated in the ER and colocalized with the fungal proteasome. These results provide an indication that there is a limit to how far T. reesei Rut-C30, already under secretion stress, can be pressed to produce higher protein yields.

An integrative framework of supervisor responses to workgroup conflict

It is well-established that prolonged exposure to workplace conflict, as a work stressor, is linked to physical illness and psychological dysfunction in employees (see Spector and Jex, 1998; Romanov, Appelberg, Honkasalo, and Koskenvuo, 1996; Skogstad, Einarsen, Torsheim, Aasland, and Hetland, 2007). In addition to the negative implications for physiological and psychological health, workplace conflict has been shown to influence employee behaviors that have consequences for organizational effectiveness (e.g., turnover and impaired performance; see Bowling and Beehr, 2006; De Dreu and Weingart, 2003). Further, research suggests that managers spend approximately 20 percent of their time managing conflict (Thomas, 1992; Baron, 1989). There also are substantial financial implications associated with workplace conflict. For example, in the United Kingdom, costs at the national level for sickness absence and replacement costs has been estimated to be close to £2 billion per annum (Hoel, Sparks, and Cooper, 2001).

A securities market operator's use of the "please explain" price query and its impact on compliance

Market operators in New Zealand and Australia, such as the New Zealand Exchange (NZX) and the Australian Securities Exchange (ASX), have the regulatory power in their listing rules to issue queries to their market participants to explain unusual fluctuations in trading price and/or volume in the market. The operator will issue a price query where it believes that the market has not been fully informed as to price relevant information. Responsive regulation theory has informed much of the regulatory debate in securities laws in the region. Price queries map onto the lower level of the enforcement pyramid envisaged by responsive regulation and are one strategy that a market operator can use in communicating its compliance expectations to its stakeholders. The issue of a price query may be a precursor to more severe enforcement activities. The aim of this study is to investigate whether increased use of price queries by the securities market operator in New Zealand corresponded with an increase in disclosure frequency by all participating companies. The study finds that an increased use of price queries did correspond with an increase in disclosure frequency. A possible explanation for this finding is that price queries are an effective means of appealing to the factors that motivate corporations, and the individuals who control them, to comply with the law and regulatory requirements. This finding will have implications for both the NZX and the ASX as well as for regulators and policy makers generally.

Elevated cyclic AMP suppresses ConA-induced MT1-MMP expression in MDA- MB-231 human breast cancer cells

We have previously reported that induction of MMP-2 activation by Concanavalin A (ConA) in MDA-MB-231 human breast cancer cells involves both transcriptional and post-transcriptional mechanisms, and that the continuous presence of ConA is required for MMP-2 activation (Yu et al. Cancer Res, 55, 3272-7, 1995). In an effort to identify signal transduction pathways which may either contribute to or modulate this mechanism, we found that three different cAMP-inducing agents, cholera toxin (CT), forskolin (FSK), and 3- isobutyl-1-methylxanthine (IBMX) partially inhibited ConA-induced MT1-MMP expression and MMP-2 activation in MDA-MB-231 cells. Combinations of CT or FSK with IBMX exhibited additive effects on reduction of MT1-MMP mRNA expression and MMP-2 activation. Agents which increase cAMP levels appeared to target transcriptional aspects of ConA induction, reducing MT1-MMP mRNA and protein in parallel with the reduced MMP-2 activation. In the absence of ConA, down-regulation of constitutive production of MT1-MMP mRNA and protein was observed, indicating that cAMP acts independently of ConA. These observations may help to elucidate factors regulating MT1-MMP expression, which may be pivotal to the elaboration of invasive machinery on the cell surface.

Vimentin and epithelial-mesenchymal transition in human breast cancer : observations in vitro and in vivo

Breast cancer is a highly prevalent disease among women worldwide. While the expression of certain proteins within these tumours is used for prognosis and selection of therapies, there is a continuing need for additional markers to be identified. A considerable amount of current literature, based predominantly on cell culture systems, suggests that a major mechanism responsible for the progression of breast cancer is due to tumour cells losing their epithelial features and gaining mesenchymal properties. These events are proposed to be very similar to the epithelial-mesenchymal transition (EMT) process that has been well characterised in embryonic development. For the developmental and putative cancer EMT, the cell intermediate filament status changes from a keratin-rich network which connects to adherens junctions and hemidesmosomes, to a vimentin-rich network connecting to focal adhesions. This review summarises observations of vimentin expression in breast cancer model systems, and discusses the potential role of EMT in human breast cancer progression, and the prognostic usefulness of vimentin expression.

The ENHANCES study--Enhancing Head and Neck Cancer patients' Experiences of Survivorship: study protocol for a randomized controlled trial

Background Few cancers pose greater challenges than head and neck (H&N) cancer. Residual effects following treatment include body image changes, pain, fatigue and difficulties with appetite, swallowing and speech. Depression is a common comorbidity. There is limited evidence about ways to assist patients to achieve optimal adjustment after completion of treatment. In this study, we aim to examine the effectiveness and feasibility of a model of survivorship care to improve the quality of life of patients who have completed treatment for H&N cancer. Methods This is a preliminary study in which 120 patients will be recruited. A prospective randomised controlled trial of the H&N Cancer Survivor Self-management Care Plan (HNCP) involving pre- and post-intervention assessments will be used. Consecutive patients who have completed a defined treatment protocol for H&N cancer will be recruited from two large cancer services and randomly allocated to one of three study arms: (1) usual care, (2) information in the form of a written resource or (3) the HNCP delivered by an oncology nurse who has participated in manual-based training and skill development in patient self-management support. The trained nurses will meet patients in a face-to-face interview lasting up to 60 minutes to develop an individualised HNCP, based on principles of chronic disease self-management. Participants will be assessed at baseline, 3 and 6 months. The primary outcome measure is quality of life. The secondary outcome measures include mood, self-efficacy and health-care utilisation. The feasibility of implementing this intervention in routine clinical care will be assessed through semistructured interviews with participating nurses, managers and administrators. Interviews with patients who received the HNCP will explore their perceptions of the HNCP, including factors that assisted them in achieving behavioural change. Discussion In this study, we aim to improve the quality of life of a patient population with unique needs by means of a tailored self-management care plan developed upon completion of treatment. Delivery of the intervention by trained oncology nurses is likely to be acceptable to patients and, if successful, will be a model of care that can be implemented for diverse patient populations.

Relating Across Difference : A Case Study in Transformative Learning

This chapter examines the process of “transformative learning” for the 2008 cohort of the St. Thomas More College–Intercordia Canada (STM/IC) international community service-learning program. My primary data comes from the eight students who were part of that cohort, when I was their program coordinator. That data later became the heart of my Master’s thesis and was approved by the Research Ethics Office of the University of Saskatchewan. In this chapter I focus on three of those eight participants, outlining the critical elements of their experiences that were conducive to their transformative learning. To be sure, my sample size is too small to draw generalizable conclusions, but the quality of information I received from these students and their colleagues, coupled with the follow-up conversations I had with other educators and with the accompanying literature, supports the value of reflecting at length here about these students’ comments. I have chosen to highlight the experiences of these three participants because together they provide a range of experiences that best enables an analysis of the conditions that both did and did not lead to transformative learning. This case study suggests that transformative learning occurs through: the dynamics of vulnerability, a discovery of persisting differences within inter-personal relationships, and an experience of welcome and hospitality in the host environment. In contrast to other studies that focus on the enhanced capacities, skills and subsequent employability of participants through international education, transformative learning for these students required a relinquishing of securities, a disorientation and critical interrogation of the self, and enhanced receptivity to the newly recognized Other. Moreover, consistent with the critical-humanistic educational philosophy shared in different ways by the participants of the STM/IC program, this study suggests that international community service-learning can most responsibly contribute to good global citizenship through the construction of relationships of solidarity across difference.

Generalised cumulative arrays in secret sharing

Cumulative arrays have played an important role in the early development of the secret sharing theory. They have not been subject to extensive study so far, as the secret sharing schemes built on them generally result in much larger sizes of shares, when compared with other conventional approaches. Recent works in threshold cryptography show that cumulative arrays may be the appropriate building blocks in non-homomorphic threshold cryptosystems where the conventional secret sharing methods are generally of no use. In this paper we study several extensions of cumulative arrays and show that some of these extensions significantly improve the performance of conventional cumulative arrays. In particular, we derive bounds on generalised cumulative arrays and show that the constructions based on perfect hash families are asymptotically optimal. We also introduce the concept of ramp perfect hash families as a generalisation of perfect hash families for the study of ramp secret sharing schemes and ramp cumulative arrays.