Comparison of the modes of action of Apremilast and Roflumilast

The phosphodiesterase 4 (PDE4) family are cAMP specific phosphodiesterases that play an important role in the inflammatory response and is the major PDE type found in inflammatory cells. A significant number of PDE4 specific inhibitors have been developed and are currently being investigated for use as therapeutic agents. Apremilast, a small molecule inhibitor of PDE 4 is in development for chronic inflammatory disorders and has shown promise for the treatment of psoriasis, psoriatic arthritis as well as other inflammatory diseases. It has been found to be safe and well tolerated in humans and in March 2014 it was approved by the US food and drug administration for the treatment of adult patients with active psoriatic arthritis. The only other PDE4 inhibitor on the market is Roflumilast and it is used for treatment of respiratory disease. Roflumilast is approved in the EU for the treatment of COPD and was recently approved in the US for treatment to reduce the risk of COPD exacerbations. Roflumilast is also a selective PDE4 inhibitor, administered as an oral tablet once daily, and is thought to act by increasing cAMP within lung cells. As both (Apremilast and Roflumilast) compounds selectively inhibit PDE4 but are targeted at different diseases, there is a need for a clear understanding of their mechanism of action (MOA). Differences and similarity of MOA should be defined for the purposes of labelling, for communication to the scientific community, physicians, and patients, and for an extension of utility to other diseases and therapeutic areas. In order to obtain a complete comparative picture of the MOA of both inhibitors, additional molecular and cellular biology studies are required to more fully elucidate the signalling mediators downstream of PDE4 inhibition which result in alterations in pro- and anti-inflammatory gene expression. My studies were conducted to directly compare Apremilast with Roflumilast, in order to substantiate the differences observed in the molecular and cellular effects of these compounds, and to search for other possible differentiating effects. Therefore the main aim of this thesis was to utilise cutting-edge biochemical techniques to discover whether Apremilast and Roflumilast work with different modes of action. In the first part of my thesis I used novel genetically encoded FRET based cAMP sensors targeted to different intracellular compartments, in order to monitor cAMP levels within specific microdomains of cells as a consequence of challenge with Apremilast and Roflumilast, which revealed that Apremilast and Roflumilast do regulate different pools of cAMP in cells. In the second part of my thesis I focussed on assessing whether Apremilast and Roflumilast cause differential effects on the PKA phosphorylation state of proteins in cells. I used various biochemical techniques (Western blotting, Substrate kinase arrays and Reverse Phase Protein array and found that Apremilast and Roflumilast do lead to differential PKA substrate phosphorylation. For example I found that Apremilast increases the phosphorylation of Ribosomal Protein S6 at Ser240/244 and Fyn Y530 in the S6 Ribosomal pathway of Rheumatoid Arthritis Synovial fibroblast and HEK293 cells, whereas Roflumilast does not. This data suggests that Apremilast has distinct biological effects from that of Roflumilast and could represent a new therapeutic role for Apremilast in other diseases. In the final part of my thesis, Phage display technology was employed in order to identify any novel binding motifs that associate with PDE4 and to identify sequences that were differentially regulated by the inhibitors in an attempt to find binding motifs that may exist in previously characterised signalling proteins. Petide array technology was then used to confirm binding of specific peptide sequences or motifs. Results showed that Apremilast and Roflumilast can either enhance or decrease the binding of PDE4A4 to specific peptide sequences or motifs that are found in a variety of proteins in the human proteome, most interestingly Ubiquitin-related proteins. The data from this chapter is preliminary but may be used in the discovery of novel binding partners for PDE4 or to provide a new role for PDE inhibition in disease. Therefore the work in this thesis provides a unique snapshot of the complexity of the cAMP signalling system and is the first to directly compare action of the two approved PDE4 inhibitors in a detailed way.

Quasinormal modes of black holes in anti-de Sitter space: A numerical study of the eikonal limit

Using series solutions and time-domain evolutions, we probe the eikonal limit of the gravitational and scalar-field quasinormal modes of large black holes and black branes in anti-de Sitter backgrounds. These results are particularly relevant for the AdS/CFT correspondence, since the eikonal regime is characterized by the existence of long-lived modes which (presumably) dominate the decay time scale of the perturbations. We confirm all the main qualitative features of these slowly damped modes as predicted by Festuccia and Liu [G. Festuccia and H. Liu, arXiv:0811.1033.] for the scalar-field (tensor-type gravitational) fluctuations. However, quantitatively we find dimensional-dependent correction factors. We also investigate the dependence of the quasinormal mode frequencies on the horizon radius of the black hole (brane) and the angular momentum (wave number) of vector- and scalar-type gravitational perturbations.

Localized modes of binary mixtures of Bose-Einstein condensates in nonlinear optical lattices

The properties of the localized states of a two-component Bose-Einstein condensate confined in a nonlinear periodic potential (nonlinear optical lattice) are investigated. We discuss the existence of different types of solitons and study their stability by means of analytical and numerical approaches. The symmetry properties of the localized states with respect to nonlinear optical lattices are also investigated. We show that nonlinear optical lattices allow the existence of bright soliton modes with equal symmetry in both components and bright localized modes of mixed symmetry type, as well as dark-bright bound states and bright modes on periodic backgrounds. In spite of the quasi-one-dimensional nature of the problem, the fundamental symmetric localized modes undergo a delocalizing transition when the strength of the nonlinear optical lattice is varied. This transition is associated with the existence of an unstable solution, which exhibits a shrinking (decaying) behavior for slightly overcritical (undercritical) variations in the number of atoms.

Stability, causality, and quasinormal modes of cosmic strings and cylinders

In this work we consider the evolution of a massive scalar field in cylindrically symmetric space-times. Quasinormal modes have been calculated for static and rotating cosmic cylinders. We found unstable modes in some cases. Rotating as well as static cosmic strings, i.e., without regular interior solutions, do not display quasinormal oscillation modes. We conclude that rotating cosmic cylinder space-times that present closed timelike curves are unstable against scalar perturbations.

Quasinormal modes of brane-localized standard model fields in Gauss-Bonnet theory

We study the massless scalar, Dirac, and electromagnetic fields propagating on a 4D-brane, which is embedded in higher-dimensional Gauss-Bonnet space-time. We calculate, in the time domain, the fundamental quasinormal modes of a spherically symmetric black hole for such fields. Using WKB approximation we study quasinormal modes in the large multipole limit. We observe also a universal behavior, independent on a field and value of the Gauss-Bonnet parameter, at an asymptotically late time.

Gravitational quasinormal modes of AdS black branes in d spacetime dimensions

The AdS/CFT duality has established a mapping between quantities in the bulk AdS black-hole physics and observables in a boundary finite-temperature field theory. Such a relationship appears to be valid for an arbitrary number of spacetime dimensions, extrapolating the original formulations of Maldacena`s correspondence. In the same sense properties like the hydrodynamic behavior of AdS black-hole fluctuations have been proved to be universal. We investigate in this work the complete quasinormal spectra of gravitational perturbations of d-dimensional plane-symmetric AdS black holes (black branes). Holographically the frequencies of the quasinormal modes correspond to the poles of two-point correlation functions of the field-theory stress-energy tensor. The important issue of the correct boundary condition to be imposed on the gauge-invariant perturbation fields at the AdS boundary is studied and elucidated in a fully d-dimensional context. We obtain the dispersion relations of the first few modes in the low-, intermediate- and high-wavenumber regimes. The sound-wave (shear-mode) behavior of scalar (vector)-type low- frequency quasinormal mode is analytically and numerically confirmed. These results are found employing both a power series method and a direct numerical integration scheme.

Evaluation of proximal caries in images resulting from different modes of radiographic digitalization

Objective: The aim of this study was to evaluate the performances of observers in diagnosing proximal caries in digital images obtained from digital bitewing radiographs using two scanners and four digital cameras in Joint Photographic Experts Group (JPEG) and tagged image file format (TIFF) files, and comparing them with the original conventional radiographs. Method: In total, 56 extracted teeth were radiographed with Kodak Insight film (Eastman Kodak, Rochester, NY) in a Kaycor Yoshida X-ray device (Kaycor X-707;Yoshida Dental Manufacturing Co., Tokyo, Japan) operating at 70 kV and 7 mA with an exposure time of 0.40 s. The radiographs were obtained and scanned by CanonScan D646U (Canon USA Inc., Newport News, VA) and Genius ColorPage HR7X (KYE Systems Corp. America, Doral, FL) scanners, and by Canon Powershot G2 (Canon USA Inc.), Canon RebelXT (Canon USA Inc.), Nikon Coolpix 8700 (Nikon Inc., Melville, NY), and Nikon D70s (Nikon Inc.) digital cameras in JPEG and TIFF formats. Three observers evaluated the images. The teeth were then observed under the microscope in polarized light for the verification of the presence and depth of the carious lesions. Results: The probability of no diagnosis ranged from 1.34% (Insight film) to 52.83% (CanonScan/JPEG). The sensitivity ranged from 0.24 (Canon RebelXT/JPEG) to 0.53 (Insight film), the specificity ranged from 0.93 (Nikon Coolpix/JPEG, Canon Powershot/TIFF, Canon RebelXT/JPEG and TIFF) to 0.97 (CanonScan/TIFF and JPEG) and the accuracy ranged from 0.82 (Canon RebelXT/JPEG) to 0.91 (CanonScan/JPEG). Conclusion: The carious lesion diagnosis did not change in either of the file formats (JPEG and TIFF) in which the images were saved for any of the equipment used. Only the CanonScan scanner did not have adequate performance in radiography digitalization for caries diagnosis and it is not recommended for this purpose. Dentomaxillofacial Radiology (2011) 40, 338-343. doi: 10.1259/dmfr/67185962

Reliability and Failure Modes of Implant-Supported Y-TZP and MCR Three-Unit Bridges

Purpose: Chipping within veneering porcelain has resulted in high clinical failure rates for implant-supported zirconia (yttria-tetragonal zirconia polycrystals [Y-TZP]) bridges. This study evaluated the reliability and failure modes of mouth-motion step-stress fatigued implant-supported Y-TZP versus palladium-silver alloy (PdAg) three-unit bridges. Materials and Methods: Implant-abutment replicas were embedded in polymethylmethacrylate resin. Y-TZP and PdAg frameworks, of similar design (n = 21 each), were fabricated, veneered, cemented (n = 3 each), and Hertzian contact-tested to obtain ultimate failure load. In each framework group, 18 specimens were distributed across three step-stress profiles and mouth-motion cyclically loaded according to the profile on the lingual slope of the buccal cusp of the pontic. Results: PdAg failures included competing flexural cracking at abutment and/or connector area and chipping, whereas Y-TZP presented predominantly cohesive failure within veneering porcelain. Including all failure modes, the reliability (two-sided at 90% confidence intervals) for a ""mission"" of 50,000 and 100,000 cycles at 300 N load was determined (Alta Pro, Reliasoft, Tucson, AZ, USA). No difference in reliability was observed between groups for a mission of 50,000. Reliability remained unchanged for a mission of 100,000 for PdAg, but significantly decreased for Y-TZP. Conclusions: Higher reliability was found for PdAg for a mission of 100,000 cycles at 300 N. Failure modes differed between materials.

Fatigue life and failure modes of crowns systems with a modified framework design

Objectives: To evaluate the effect of framework design on the fatigue life and failure modes of metal ceramic (MC, Ni-Cr alloy core, VMK 95 porcelain veneer), glass-infiltrated alumina (ICA, In-Ceram Alumina/VM7), and veneered yttria-stabilized tetragonal zirconia polycrystals (Y-TZP, IPSe.max ZirCAD/IPS e.max,) crowns. Methods: Sixty composite resin tooth replicas of a prepared maxillary first molar were produced to receive crowns systems of a standard (MCs, ICAs, and Y-TZPs, n = 10 each) or a modified framework design (MCm, ICAm, and Y-TZPm, n = 10 each). Fatigue loading was delivered with a spherical steel indenter (3.18 mm radius) on the center of the occlusal surface using r-ratio fatigue (30-300 N) until completion of 10(6) cycles or failure. Fatigue was interrupted every 125,000 cycles for damage evaluation. Weibull distribution fits and contour plots were used for examining differences between groups. Failure mode was evaluated by light polarized and SEM microscopy. Results: Weibull analysis showed the highest fatigue life for MC crowns regardless of framework design. No significant difference (confidence bound overlaps) was observed between ICA and Y-TZP with or without framework design modification. Y-TZPm crowns presented fatigue life in the range of MC crowns. No porcelain veneer fracture was observed in the MC groups, whereas ICAs presented bulk fracture and ICAm failed mainly through the veneer. Y-TZP crowns failed through chipping within the veneer, without core fractures. Conclusions: Framework design modification did not improve the fatigue life of the crown systems investigated. Y-TZPm crowns showed comparable fatigue life to MC groups. Failure mode varied according to crown system. (C) 2010 Elsevier Ltd. All rights reserved.

Failure modes of Y-TZP crowns at different cusp inclines

Objectives To compare the reliability of the disto-facial (DF) and mesio-lingual (ML) cusps of an anatomically correct zirconia (Y-TZP) crown system The research hypotheses tested were (1) fatigue reliability and failure mode are similar for the ML and DF cusps, (2) failure mode of one cusp does not affect the failure of the other Methods The average dimensions of a mandibular first molar crown were imported into CAD software, a tooth preparation was modelled by 1 5 mm marginal high reduction of proximal walls and occlusal surface by 2 0 mm The CAD-based tooth preparation was milled and used as a die to fabricate crowns (n = 14) with porcelain veneer on a 0 5 mm Y-TZP core. Crowns were cemented on composite reproductions of the tooth preparation The crowns were step-stress mouth motion fatigued with sliding (0 7 mm) a tungsten-carbide indenter of 6 25 mm diameter down on the inclines of either the DF or ML cusps Use level probability Weibull curve with use stress of 200 N and the reliability for completion of a mission of 50,000 cycles at 200 N load were calculated Results Reliability for a 200 N at 50,000 cycles mission was not different between tested cusps SEM imaging showed large cohesive failures within the veneer for the ML and smaller for the DF Fractures originated from the contact area regardless of the cusp loaded Conclusion No significant difference on fatigue reliability was observed between the DF compared to the ML cusp Fracture of one cusp did not affect the other (c) 2010 Elsevier Ltd All rights reserved

Ultra-high-temperature (UHT) treatment of milk: comparison of direct and indirect modes of heating

UHT processing of milk and its subsequent storage causes several changes which affect the shelf-life of UHT milk although it remains 'commercially sterile'. These changes include whey protein denaturation, protein-protein interaction, lactose-protein interaction, isomerisation of lactose, Maillard browning, sulphydryl compound formation, formation of a range of carbonyl and other flavoursome compounds, and formation of insoluble substances. They ultimately reduce the quality and limit the shelf life of UHT milk through development of off-flavours, fat separation, age gelation and sedimentation. The extent of these changes depends on many factors, a major one being the type of UHT heating. This review compares the effect heating milk by direct and indirect modes on various aspects of processing and quality of UHT milk.

Differential modes of alaryngeal communication and long-term voice outcomes following pharyngolaryngectomy and laryngectomy

Patterns of vocal rehabilitation for 37 pharyngolaryngectomy patients and 55 total laryngectomy patients over a 5-year period were compared. An electrolarynx (EL) was introduced as the initial communication mode immediately after surgery for 98% of patients, with 30% of pharyngolaryngectomy and 74% of laryngectomy patients subsequently developing tracheoesophageal speech (TES) as their primary mode of communication. Follow-up with 14 of 37 pharyngolaryngectomy patients and 36 of 55 laryngectomy patients was conducted 1-6 years following surgery and revealed that 90% of the pharyngolaryngectomy patients maintained the use of TES in the long term compared to 69% of the laryngectomy group. Long-term outcomes relating to communication disability and handicap did not differ significantly between the two surgical groups, however the laryngectomy patients had significantly higher levels of wellbeing. Across the whole group of patients, statistical comparison revealed that patients using TES had significantly lower levels of disability, handicap and distress than EL users. Considering that lower levels of disability, handicap and distress are associated with TES, and the data supports that suitably selected patients can maintain functional TES in the long term, increased application of this form of communication rehabilitation should be encouraged where viable for the pharyngolaryngectomy population. Copyright (C) 2003 S. Karger AG, Basel.

Trends in the modes of delivery and their impact on perinatal mortality rates

OBJECTIVE: To determine changes in the incidence of vaginal deliveries, cesarean sections, and forceps deliveries and their potential association with fetal, early neonatal, and perinatal mortality rates over time. METHODS: A retrospective study was carried out and the occurrence of deliveries supervised by university services between January 1991 and December 2000 was determined. Data regarding fetal, early neonatal, and perinatal deaths were assessed using obstetric and pediatric records and autopsy reports. RESULTS: Of a total of 33,360 deliveries, the incidence of vaginal deliveries, cesarean sections, and forceps deliveries was relatively steady (around 60, 30, and 10%, respectively) while, at the same time, there was a marked reduction in fetal mortality (from 33.3 to 13.0‰), early neonatal mortality (from 30.6 to 9.0‰), and perinatal mortality (from 56.4 to 19.3‰). CONCLUSIONS: The marked reduction in perinatal mortality rates seen during the study period without an increase in cesarean sections indicates that the decrease in perinatal mortality was not impacted by cesarean section rates. The plausible hypothesis seems to be that the reduction in perinatal mortality of deliveries performed under the supervision of university services was more likely to be associated with better neonatal care rather than the mode of delivery.