The Kinematics of interference mechanisms in certain machining operations

The mechanism of "Helical Interference" in milled slots is examined and a coherent theory for the geometry of such surfaces is presented. An examination of the relevant literature shows a fragmented approach to the problem owing to its normally destructive nature, so a complete analysis is developed for slots of constant lead, thus giving a united and exact theory for many different setting parameters and a range of cutter shapes. For the first time, a theory is developed to explain the "Interference Surface" generated in variable lead slots for cylindrical work and attention is drawn to other practical surfaces, such as cones, where variable leads are encountered. Although generally outside the scope of this work, an introductory analysis of these cases is considered in order to develop the cylindrical theory. Special emphasis is laid upon practical areas where the interference mechanism can be used constructively and its application as the rake face of a cutting tool is discussed. A theory of rake angle for such cutting tools is given for commonly used planes, and relative variations in calculated rake angle between planes is examined. Practical tests are conducted to validate both constant lead and variable lead theories and some design improvements to the conventional dividing head are suggested in order to manufacture variable lead workpieces, by use of a "superposed" rotation. A prototype machine is manufactured and its kinematic principle given for both linear and non-linearly varying superposed rotations. Practical workpieces of the former type are manufactured and compared with analytical predictions,while theoretical curves are generated for non-linear workpieces and then compared with those of linear geometry. Finally suggestions are made for the application of these principles to the manufacture of spiral bevel gears, using the "Interference Surface" along a cone as the tooth form.

Prestressed concrete beams with stirrups subjected to torsion, bending and shear

Reported in this thesis are test results of 37 eccentrically prestressed beams with stirrups. Single variable parameters were investigated including the prestressing force, the prestressing steel area, the concrete strength, the aspect ratio h/b and the stirrups size and spacing. Interaction of bending, torsion and shear was also investigated by testing a series of beams subjected to varying bending/torsional moment ratios. For the torsional strength an empirical expression of linear format is proposed and can be rearranged in a non-dimensional interaction form: T/To+V/Vo+M/Mo+Ps/Po+Fs/Fo=Pc2/Fsp. This formula which is based on an average experimental steel stress lower than the yield point is compared with 243 prestressed beams containing ' stirrups, including the author's test beams, and good agreement is obtained. For the theoretical analysis of the problem of torsion combined with bending and shear in concrete beams with stirrups, the method of torque-friction is proposed and developed using an average steel stress. A general linear interaction equation for combined torsion with bending and/or shear is proposed in the following format: (fi) T/Tu=1 where (fi) is a combined loading factor to modify the pure ultimate strength for differing cases of torsion with bending and/or shear. From the analysis of 282 reinforced and prestressed concrete beams containing stirrups, including the present investigation, good agreement is obtained between the method and the test results. It is concluded that the proposed method provides a rational and simple basis for predicting the ultimate torisional strength and may also be developed for design purposes.

Improved production of industrially relevant recombinant proteins: application of multi-factorial design of experiments and scalable process modelling

The work described in this thesis focuses on the use of a design-of-experiments approach in a multi-well mini-bioreactor to enable the rapid establishments of high yielding production phase conditions in yeast, which is an increasingly popular host system in both academic and industrial laboratories. Using green fluorescent protein secreted from the yeast, Pichia pastoris, a scalable predictive model of protein yield per cell was derived from 13 sets of conditions each with three factors (temperature, pH and dissolved oxygen) at 3 levels and was directly transferable to a 7 L bioreactor. This was in clear contrast to the situation in shake flasks, where the process parameters cannot be tightly controlled. By further optimisating both the accumulation of cell density in batch and improving the fed-batch induction regime, additional yield improvement was found to be additive to the per cell yield of the model. A separate study also demonstrated that improving biomass improved product yield in a second yeast species, Saccharomyces cerevisiae. Investigations of cell wall hydrophobicity in high cell density P. pastoris cultures indicated that cell wall hydrophobin (protein) compositional changes with growth phase becoming more hydrophobic in log growth than in lag or stationary phases. This is possibly due to an increased occurrence of proteins associated with cell division. Finally, the modelling approach was validated in mammalian cells, showing its flexibility and robustness. In summary, the strategy presented in this thesis has the benefit of reducing process development time in recombinant protein production, directly from bench to bioreactor.

Design, synthesis and biological evaluation of potential antibacterial oligonucleotides

Purine and pyrimidine triplex-forming oligonucleotides (TFOs), as potential antibacterial agents, were designed to bind by Hoogsteen and reverse Hoogsteen hydrogen bonds in a sequence specific manner in the major groove of genomic DNA at specific polypurine sites within the gyrA gene of E. coli and S. pneumoniae. Sequences were prepared by automated synthesis, with purification and characterisation determined by high performance liquid chromatograpy, capillary electrophoresis and mass spectrometry. Triplex stability was assessed using melting curves where the binding of the third strand to the duplex target, was assessed over a temperature range of 0-80°C, and at pH 6.4 and 7.2. The most successful of the unmodified TFOs (6) showed a Tm value of 26 °C at both pH values with binding via reverse Hoogsteen bonds. Binding to genomic DNA was also demonstrated by spectrofluorimetry, using fluorescein-labelled TFOs, from which dissociation constants were determined. Modifications in the form of 5mC, 5' acridine attachment, phosphorothioation, 2'-0-methylation and phosphoramidation, were made in order to. increase Tm values. Phosphoramidate modification was the most with increased Tm values of 42°C. However, the final purity of these sequences was poor due to their difficult syntheses. FACS (fluorescent activated cell sorting) analysis was used to determine the potential uptake of a fluorescently labelled analogue of 6 via passive, coJd shock mediated, and anionic liposome aided, uptake. This was established at 20°C and 37°C. At both temperatures anionic lipid-mediated uptake produced unrivalled fluorescence, equivalent to 20 and 43% at 20 and 37°C respectively. Antibacterial activity of each oligonucleotide was assessed by viable count anaJysis relying on passive uptake, cold shocking techniques, chlorpromazine-mediated uptake, and, cationic and anionic lipid-aided uptake. All oligonucleotides were assessed for their ability to enhance uptake, which is a major barrier to the effectiveness of these agents. Compound 6 under cold shocking conditions produced the greatest consistent decline in colony forming units per ml. Results for this compound were sometimes variable indicating inconsistent uptake by this particular assay method.

Integrated manufacturing systems design in a multi-product company

Using a hydraulic equipment manufacturing plant as the case study, this work explores the problems of systems integration in manufacturing systems design, stressing the behavioural aspects of motivation and participation, and the constraints involved in the proper consideration of the human sub-system. The need for a simple manageable modular organisation structure is illustrated, where it is shown, by reference to systems theory, how a business can be split into semi-autonomous operating units. The theme is the development of a manufacturing system based on an analysis of the business, its market, product, technology and constraints, coupled with a critical survey of modern management literature to develop an integrated systems design to suit a specific company in the current social environment. Society currently moves through a socio-technical revolution with man seeking higher levels of motivation. The transitory environment from an autocratic/paternalistic to a participative operating mode demands systems parameters only found to a limited extent in manufacturing systems today. It is claimed, that modern manufacturing systems design needs to be based on group working, job enrichment, delegation of decision making and reduced job monotony. The analysis shows how negative aspects of cellular manufacture such as lack of flexibility and poor fixed asset utilisation are relatively irrelevant and misleading in the broader context of the need to come to terms with the social stresses imposed on a company operating in the industrial environment of the present and the immediate future.

Design of a novel solar thermal collector using a multi-criteria decision-making methodology

Three novel solar thermal collector concepts derived from the Linear Fresnel Reflector (LFR) are developed and evaluated through a multi-criteria decision-making methodology, comprising the following techniques: Quality Function Deployment (QFD), the Analytical Hierarchy Process (AHP) and the Pugh selection matrix. Criteria are specified by technical and customer requirements gathered from Gujarat, India. The concepts are compared to a standard LFR for reference, and as a result, a novel 'Elevation Linear Fresnel Reflector' (ELFR) concept using elevating mirrors is selected. A detailed version of this concept is proposed and compared against two standard LFR configurations, one using constant and the other using variable horizontal mirror spacing. Annual performance is analysed for a typical meteorological year. Financial assessment is made through the construction of a prototype. The novel LFR has an annual optical efficiency of 49% and increases exergy by 13-23%. Operational hours above a target temperature of 300 C are increased by 9-24%. A 17% reduction in land usage is also achievable. However, the ELFR suffers from additional complexity and a 16-28% increase in capital cost. It is concluded that this novel design is particularly promising for industrial applications and locations with restricted land availability or high land costs. The decision analysis methodology adopted is considered to have a wider potential for applications in the fields of renewable energy and sustainable design. © 2013 Elsevier Ltd. All rights reserved.

Optimized design of high power mid-infrared Er3+, Pr3+ -codoped ZBLAN fiber laser

Based on the rate equations describing the operation of the Er3+, Pr3+ -codoped ZBLAN fiber lasers with different pump configurations, theoretical calculations that relate to the population characteristics and optimization of CW operation of high power Er3+, Pr3+ :ZBLAN double-clad fiber lasers are presented. Using the measured ET (energy-transfer), ETU (energy-transfer-upconversion) and CR (cross-relaxation) parameters relevant to Er3+, Pr3+ -codoped ZBLAN, a good agreement between the theoretical results from the model and recently reported experimental measurements is obtained. The effects on the slope efficiency of a number of laser parameters including fiber length, reflectance of the output mirror and pumping configuration are quantitatively analyzed and used for the design and optimization of high power Er3+, Pr3+ -codoped ZBLAN fiber lasers.

Design and development of cationic liposomes as DNA vaccine adjuvants

Cationic liposomes have been extensively explored for their efficacy in delivering nucleic acids, by offering the ability to protect plasmid DNA against degradation, promote gene expression and, in the case of DNA vaccines, induce both humoural and cellular immune responses. DNA vaccines may also offer advantages in terms of safety, but they are less effective and need an adjuvant to enhance their immunogenicity. Therefore, cationic liposomes can be utilised as delivery systems and/or adjuvants for DNA vaccines to stimulate stronger immune responses. To explore the role of liposomal systems within plasmid DNA delivery, parameters such as the effect of lipid composition, method of liposome preparation and presence of electrolytes in the formulation were investigated in characterisation studies, in vitro transfection studies and in vivo biodistribution and immunisation studies. Liposomes composed of 1,2-dioleoyl-sn-glycero 3-phosphoethanolamine (DOPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3- trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method and hydrated in aqueous media with or without presence of electrolytes. Whilst the in vitro transfection efficiency of all liposomes resulted to be higher than Lipofectin, DSTAP-based liposomes showed significantly higher transfection efficiency than DOTAP-based formulations. Furthermore, upon intramuscular injection of liposomal DNA vaccines, DSTAP-based liposomes showed a significantly stronger depot effect at the injection site. This could explain the result of heterologous immunisation studies, which revealed DSTAP-based liposomal vaccines induce stronger immune responses compared to DOTAP-based formulations. Previous studies have shown that having more liposomally associated antigen at the injection site would lead to more drainage of them into the local lymph nodes. Consequently, this would lead to more antigens being presented to antigen presenting cells, which are circulating in lymph nodes, and this would initiate a stronger immune response. Finally, in a comparative study, liposomes composed of dimethyldioctadecylammonium bromide (DDA) in combination with DOPE or immunostimulatory molecule of trehalose 6,6-dibehenate (TDB) were prepared and investigated in vitro and in vivo. Results showed that although DDA:TDB is not able to transfect the cells efficiently in vitro, this formulation induces stronger immunity compared to DDA:DOPE due to the immunostimulatory effects of TDB. This study demonstrated, while the presence of electrolytes did not improve immune responses, small unilamellar vesicle (SUV) liposomes induced stronger humoural immune responses compared to dehydration rehydration vesicle (DRV) liposomes. Moreover, lipid composition was shown to play a key role in in vitro and in vivo behaviour of the formulations, as saturated cationic lipids provided stronger immune responses compared to unsaturated lipids. Finally, heterologous prime/boost immunisation promoted significantly stronger immune responses compared to homologous vaccination of DNA vaccines, however, a single immunisation of subunit vaccine provoked comparable levels of immune response to the heterologous regimen, suggesting more immune efficiency for subunit vaccines compared to DNA vaccines.

The accuracy of parameter estimation from noisy data, with application to resonance peak estimation in distributed Brillouin sensing

Distributed Brillouin sensing of strain and temperature works by making spatially resolved measurements of the position of the measurand-dependent extremum of the resonance curve associated with the scattering process in the weakly nonlinear regime. Typically, measurements of backscattered Stokes intensity (the dependent variable) are made at a number of predetermined fixed frequencies covering the design measurand range of the apparatus and combined to yield an estimate of the position of the extremum. The measurand can then be found because its relationship to the position of the extremum is assumed known. We present analytical expressions relating the relative error in the extremum position to experimental errors in the dependent variable. This is done for two cases: (i) a simple non-parametric estimate of the mean based on moments and (ii) the case in which a least squares technique is used to fit a Lorentzian to the data. The question of statistical bias in the estimates is discussed and in the second case we go further and present for the first time a general method by which the probability density function (PDF) of errors in the fitted parameters can be obtained in closed form in terms of the PDFs of the errors in the noisy data.

Optimized design of high power mid-infrared Er3+, Pr3+ -codoped ZBLAN fiber laser

Based on the rate equations describing the operation of the Er3+, Pr3+ -codoped ZBLAN fiber lasers with different pump configurations, theoretical calculations that relate to the population characteristics and optimization of CW operation of high power Er3+, Pr3+ :ZBLAN double-clad fiber lasers are presented. Using the measured ET (energy-transfer), ETU (energy-transfer-upconversion) and CR (cross-relaxation) parameters relevant to Er3+, Pr3+ -codoped ZBLAN, a good agreement between the theoretical results from the model and recently reported experimental measurements is obtained. The effects on the slope efficiency of a number of laser parameters including fiber length, reflectance of the output mirror and pumping configuration are quantitatively analyzed and used for the design and optimization of high power Er3+, Pr3+ -codoped ZBLAN fiber lasers.

High-throughput manufacturing of size-tuned liposomes by a new microfluidics method using enhanced statistical tools for characterization

Microfluidics has recently emerged as a new method of manufacturing liposomes, which allows for reproducible mixing in miliseconds on the nanoliter scale. Here we investigate microfluidics-based manufacturing of liposomes. The aim of these studies was to assess the parameters in a microfluidic process by varying the total flow rate (TFR) and the flow rate ratio (FRR) of the solvent and aqueous phases. Design of experiment and multivariate data analysis were used for increased process understanding and development of predictive and correlative models. High FRR lead to the bottom-up synthesis of liposomes, with a strong correlation with vesicle size, demonstrating the ability to in-process control liposomes size; the resulting liposome size correlated with the FRR in the microfluidics process, with liposomes of 50 nm being reproducibly manufactured. Furthermore, we demonstrate the potential of a high throughput manufacturing of liposomes using microfluidics with a four-fold increase in the volumetric flow rate, maintaining liposome characteristics. The efficacy of these liposomes was demonstrated in transfection studies and was modelled using predictive modeling. Mathematical modelling identified FRR as the key variable in the microfluidic process, with the highest impact on liposome size, polydispersity and transfection efficiency. This study demonstrates microfluidics as a robust and high-throughput method for the scalable and highly reproducible manufacture of size-controlled liposomes. Furthermore, the application of statistically based process control increases understanding and allows for the generation of a design-space for controlled particle characteristics.

Design of multilevel amplitude regenerative system

We propose a scheme for multilevel (nine or more) amplitude regeneration based on a nonlinear optical loop mirror (NOLM) and demonstrate through numerical modeling its efficiency and cascadability on circular 16-, 64-, and 256- symbol constellations. We show that the amplitude noise is efficiently suppressed. The design is flexible and enables variation of the number of levels and their positioning. The scheme is compatible with phase regenerators. Also, compared to the traditional single-NOLM configuration scheme, new features, such as reduced and sign-varied power-dependent phase shift, are available. The model is simple to implement, as it requires only two couplers in addition to the traditional NOLM, and offers a vast range of optimization parameters. © 2014 Optical Society of America.

Multi-frequency segmental bio-impedance device:design, development and applications

Bio-impedance analysis (BIA) provides a rapid, non-invasive technique for body composition estimation. BIA offers a convenient alternative to standard techniques such as MRI, CT scan or DEXA scan for selected types of body composition analysis. The accuracy of BIA is limited because it is an indirect method of composition analysis. It relies on linear relationships between measured impedance and morphological parameters such as height and weight to derive estimates. To overcome these underlying limitations of BIA, a multi-frequency segmental bio-impedance device was constructed through a series of iterative enhancements and improvements of existing BIA instrumentation. Key features of the design included an easy to construct current-source and compact PCB design. The final device was trialled with 22 human volunteers and measured impedance was compared against body composition estimates obtained by DEXA scan. This enabled the development of newer techniques to make BIA predictions. To add a ‘visual aspect’ to BIA, volunteers were scanned in 3D using an inexpensive scattered light gadget (Xbox Kinect controller) and 3D volumes of their limbs were compared with BIA measurements to further improve BIA predictions. A three-stage digital filtering scheme was also implemented to enable extraction of heart-rate data from recorded bio-electrical signals. Additionally modifications have been introduced to measure change in bio-impedance with motion, this could be adapted to further improve accuracy and veracity for limb composition analysis. The findings in this thesis aim to give new direction to the prediction of body composition using BIA. The design development and refinement applied to BIA in this research programme suggest new opportunities to enhance the accuracy and clinical utility of BIA for the prediction of body composition analysis. In particular, the use of bio-impedance to predict limb volumes which would provide an additional metric for body composition measurement and help distinguish between fat and muscle content.

Design of few-mode fibers with up to 12 modes and low differential mode delay

In this paper, we investigate the design of few-mode fibers (FMFs) guiding 4 to 12 non-degenerate linearly polarized (LP) modes with low differential mode delay (DMD) over the C-band, suitable for long-haul transmission. The refractive index profile considered is composed by a graded-core with a cladding trench (GCCT). The optimization of the profile parameters aims the lowest possible DMD and macro-bend losses (MBL) lower than the ITU-T standard recommendation. The optimization results show that the optimum DMD and the MBL scale with the number of modes. Additionally, it is shown that the refractive-index relative difference at the core center is one of the most preponderant parameters, allowing to reduce the DMD at the expense of increasing MBL. Finally, the optimum DMD obtained for 12 LP modes is lower than 3 ps/km. © 2014 IEEE.

Design of few-mode fibers with M-modes and low differential mode delay

In this paper, we investigate the design of few-mode fibers (FMFs) guiding 2 to 12 linearly polarized (LP) modes with low differential mode delay (DMD) over the C-band, suitable for long-haul transmission. Two different types of refractive index profile have been considered: a graded-core with a cladding trench (GCCT) profile and a multi-step-index (MSI) profile. The profiles parameters are optimized in order to achieve: the lowest possible DMD and macro-bend losses (MBL) lower than the ITU-T standard recommendation. The optimization results show that the MSI profiles present lower DMD than the minimum achieved with a GCCT profile. Moreover, it is shown that the optimum DMD and the MBL scale with the number of modes for both profiles. The optimum DMD obtained for 12 LP modes is lower than 3 ps/km using a GCCT profile and lower than 2.5 ps/km using a MSI profile. The optimization results reveal that the most preponderant parameter of the GCCT profile is the refractive index relative difference at the core center, Δnco. Reducing Δn co, the DMD is reduced at the expense of increasing the MBL. Regarding the MSI profiles, it is shown that 64 steps are required to obtain a DMD improvement considering 12 LP modes. Finally, the impact of the fabrication margins on the optimum DMD is analyzed. The probability of having a manufactured FMF with 12 LP modes and DMD lower than 12 ps/km is approximately 68% using a GCCT profile and 16% using a MSI profile. © 2013 IEEE.