Dextran enzyme imine complexes : a preliminary study : this thesis was presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University

A model system involving the formation of protein-dextran complexes has been investigated with a view to improving existing methods of drug administration. Activation of the dextran was achieved by periodate oxidation to give levels of 7%, 21% and 56% activated glucose moieties. The protein-dextran complexes were investigated with the prospect of obtaining sustained release of proteins from the dextran in an unmodified form. Covalent conjugation of proteins to carbohydrate polymers is known to confer stability on the protein. The proteins in this study were bound to the dextran through imine bonds. The proteins investigated were lysozyme, trypsin, amylase, alcohol dehydrogenase and catalase. The selection covered a range of molecular weights and varying enzymatic activities. As might be predicted, the speed of complex formation was shown to be greater at the 21% level of activation compared to the 7% activation of dextran in all cases studied. Lysozyme, the smallest protein, readily formed complexes at all three levels of activation. At the 56% level the resulting complex had an extremely high MW, greater than 1MDa. The extensive binding between the dextran and lysozyme molecules resulted in a complex that was inactive and showed no signs of releasing any lysozyme, active or inactive. At the lower levels of activation, complex was formed with relative ease. Upon conjugation lysozyme exhibited only minimal activity. Release of a lysozyme-like species with normal lytic activity was observed. Precautions were taken to minimise possible autolysis in the trypsin study. Once complexed it was postulated that autolysis would be prevented or minimised. Similarly the 56% level of activation appeared to be too high to obtain a viable complex for facile trypsin release. Sustained release of a trypsin-like protein was observed with complexes at the 7% and 21% levels. SEC and SDS-PAGE, in conjunction with a positive BAPNA assay gave support to the released species being trypsin-like. While complexed to the dextran trypsin showed no signs of activity. Released trypsin-like species and unreacted trypsin showed similar tryptic maps from a synthetic peptide, the peptide was designed to show distinctive fragments. α-Amylase, twice the MW of trypsin and over three times the MW of lysozyme, formed complexes with ease at both 7% and 21% levels of activation. Conjugation to dextran did not effect the activity of α-amylase. Over time the release of an α-amylase-like species from the complex was observed. Alcohol dehydrogenase and catalase are both high MW proteins. Complex formation was observed for each protein. Subsequent experiments showed that upon release the proteins appeared to dissociate, most probably into their subunits. It is also possible that the dimers and monomers bound to the dextran. The main advantage of conjugation in this case appeared to be to confer stability on the proteins. The ADH-complex exhibited enzymatic activity. At 7% and 21% activation levels the lower MW proteins formed complexes with dextran that exhibited release of a protein species. The higher MW proteins were possibly stabilised when conjugated to dextran, but dissociated upon release. Investigations have shown that the level of activation chosen affects the extent of binding and therefore the functions of the resultant complex. Thus activation levels can be manipulated depending on the desired result. While lower dextran activation levels appeared to be more suited for smaller MW proteins, there were indications that the larger MW proteins could form beneficial complexes at higher activation levels. Results indicated that conjugation to periodate activated dextran could be extended to further proteins with the possibility of therapeutic or commercial applications

Including Disabled People in Sanitation and Hygiene Services

The needs of disabled people in developing countries are consistently overlooked when it comes to providing sanitation and hygiene services. This reality has severe and widespread consequences for the health, dignity, education, and employment of disabled people and their caregivers. This briefing note explores these issues and suggests how more and better research could influence policy and improve programmes

Voice of Silence: Women Inmates\u27 Perspective on Sexual Violence in the Soviet Gulag, 1936-1956

Senior Project submitted to The Division of Social Studies of Bard College

Integrating upstream and downstream process development strategies for mammalian cell derived therapeutic antibodies

Recent improvements in volumetric antibody productivity (often in excess of 5 g/L) have been achieved by advances in cell lines and upstream processing, but often lead to harvest material becoming more difficult to recover. These intensified upstream operations require a renewed prioritisation of the integration of upstream and downstream process development to ensure product purification issues are taken into consideration, to avoid extensive and expensive clearance strategies downstream. Here, it was demonstrated that changes to upstream process parameters at the bioreactor stage of monoclonal antibody production affect product quantity and quality. Culture pH, temperature and seed density setpoints leading to high titre are commonly also linked to higher post-protein A HCP levels, reduced monomer percentages and increased percentages of undesirable glycan structures. To predict post-protein A product quality, several potential indicators that can be measured in harvest material (prior to using expensive purification resources) were explored, including culture viability and osmolality, revealing unexpectedly that culture viability could not be used for such a purpose, but that osmolality has the potential to be used as a product quality indicator. The impact of culture duration on product quality was also investigated and it was shown that as cultivation progressed and antibody titre increased, product quality declined, in one case due to post-protein A HCP levels increasing by 75% from day 14 to day 17 of culture. HCP identification by mass spectrometry was applied to this system to provide insights into cellular behaviour and HCP carryover during protein A purification. It showed increases in several classes of post-protein A HCPs (e.g. stress response proteins) as the culture progressed, particularly on days 15 and 17 of culture which were associated with significant increases in total HCP levels. This provides a new level of insight into HCPs that are retained during mAb purification which may be used to aide process development strategies

Offspring of the Aurochs: A comparison of a reference collection of Dexter cattle skeletons with archaeological and historical data

This thesis aimed to re-evaluate archaeological interpretations of cattle exploitation strategies and propose new and integrated interpretations informed by archaeological and historical evidence. In addition, the study used the modern Dexter breed as a comparandum for the small cattle represented on British archaeological sites from the Bronze Age onwards. Data from a real death assemblage of forty complete and partial skeletons of Dexter cattle in the author’s reference collection, ranging in age from stillborn calves to an eighteen year old cow, and eight partial skeletons of juvenile Jersey and Jersey cross cattle were presented. The analysis was approached from the management of live cattle by age and sex groupings, rather than bones, an approach influenced by recent archaeological interpretations of the concept of the Operational Chain, or ‘biography’, incorporating the social aspect into the analysis. Critical examination and integration of information from historical and modern sources pertaining to husbandry, together with observations made in the author’s Dexter herd, were used to give insights into the reconstruction of herding strategies. The data from the Dexters, of known life history, demonstrated that simplistic hypotheses, for example equating a suite of pathological changes as evidence for draught cattle, are untenable and also suggested a new interpretation of the aetiology of osteochondrosis in young male cattle. The dwarf cattle in the Dexter sample demonstrated the difficulty of recognising bones of such animals in highly fragmented assemblages but also that metrical separation from estimated withers heights is feasible. Interpretations of “improvement” in cattle size prior to the 18th century were debated in detail. Both “profit” and the input of women in dairy-based herding were proposed as driving forces for variations in cattle size and the age and sex groupings in the slaughter population. The Dexter data were shown to be applicable to the interpretation of archaeological cattle bone assemblages from a wide range of prehistoric to later post-medieval sites in England and Wales. This thesis has demonstrated that new and nuanced interpretations of cattle bone assemblages are possible, integrating foodways and social aspects of herding

Importancia del desarrollo del aprendizaje emocional y social dentro de la clase de AICLE haciendo especial referencia a España

The Spanish Law of Education (LOMCE)1 of 2013 states that teachers and schools have the responsibility to foster students’ personal growth and social integration (BOE 2013, Article 12886, p. 97858-9). Meanwhile, the progressive implementation of bilingual education in many autonomous communities within Spain, such as the Autonomous Community of Madrid, has necessarily shifted teaching towards approaches such as Content and Language Integrated Learning (CLIL). This article presents an argument for the compatibility and mutual inclusiveness of CLIL and the objectives of the LOMCE regarding social and emotional learning.La Ley Orgánica para la Mejora de la Calidad Educativa (LOMCE) de 2013, expone que la educación debe facilitar el desarrollo personal y la integración social en los alumnos (BOE 2013, Artículo 12886, p. 97858-9). A la vez, la implantación progresiva de la educación bilingüe en muchas comunidades autónomas en España, como la de Madrid, ha hecho necesaria la inclusión de metodologías como el Aprendizaje Integrado de Contenidos y Lengua Extranjera (AICLE). Ese artículo argumenta la compatibilidad y mutua inclusividad de AICLE y los objetivos de la LOMCE en lo que respecta al desarrollo personal e integración social

Understanding sustained behavior change: the role of life crises and the process of reinvention

Much research has addressed behavior change but has tended to focus on short term changes. This study aimed to explore the mechanisms behind sustained changes in behavior and involved qualitative interviews with 34 `success stories' who had either lost weight through changes in diet and exercise (n = 24) or stopped smoking (n = 10) and had maintained this change for at least 3 years. The results showed that the majority described how their sustained behavior change had been triggered by a significant life crisis relating to their health, relationships or salient milestones. This initial change was then translated into sustained change if three sustaining conditions were met: the function of the unhealthy behavior was disrupted, the individual perceived that their choice over carrying out the unhealthy behavior had been reduced and they adhered to a behavioral model of their problem. Further, these conditions functioned by enabling a process of reinvention with participants showing a shift in identity toward a new healthier self. The results are discussed in terms of self-regulation and the establishment of a new post-crisis healthier equilibrium

A Multidisciplinary Study of Biomarkers in Hydrothermal Deposits: Applications to the Search for Life on Mars

Hydrothermal systems have been suggested as suitable environments for the appearance of life on the Early Earth and may provide habitats for microorganisms on Mars. The deposits created by these systems are preserved in the geological record. This research investigates the key mineralogical, textural and biological markers found in terrestrial hydrothermal deposits that can be used as analogues in the search for evidence of life on Mars. Samples of silica sinter from Iceland and New Zealand, the Rhynie Chert 396 Ma old deposit from Aberdeenshire and hydrothermally altered impactites from the Chicxulub impact crater have been analysed to understand the mineralogical properties unique to the different hydrothermal conditions and the evidence of extant or extinct microorganisms within them. Re-colonisation of basaltic substrates by hot spring-derived cultures was also carried out. This research was conducted using a multidisciplinary approach with the analytical techniques and instruments involved currently used for in-situ and orbital observations of planetary bodies. The principal techniques, Fourier Transform Infrared (FTIR) spectroscopy and Gas Chromatography Mass Spectrometry (GCMS), were used due to their combined capabilities in the identification of a variety of rocks and minerals, and a wide range of organic compounds. This PhD research has shown that FTIR in particular is an exceptional analytical technique for use in astrobiological investigations. This research has characterised hydrothermal deposits of different ages and created by different processes on Earth to ascertain their potential for preserving organic compounds in similar deposits on Mars. Results indicate that siliceous hydrothermal deposits of recent and ancient formation yield biomolecular evidence for past and present microbial colonisation as do hydrothermally altered impact deposits and re-colonised basaltic substrates. The identification of mineralogical and biological information using FTIR reflectance spectroscopy has wide implications in the search for life on Mars and other planetary bodies

Alvaro Moreno and Matteo Mossio. 2015. Biological autonomy: a philosophical and theoretical enquiry

Review of Alvaro Moreno and Matteo Mossio. 2015. Biological autonomy: a philosophical and theoretical enquir