Novel ingredients from brewers' spent grain - bioactivity in cell culture model systems and bioactivity retention in fortified food products

Functional food ingredients, with scientifically proven and validated bioactive effects, present an effective means of inferring physiological health benefits to consumers to reduce the risk of certain diseases. The search for novel bioactive compounds for incorporation into functional foods is particularly active, with brewers’ spent grain (BSG, a brewing industry co-product) representing a unique source of potentially bioactive compounds. The DNA protective, antioxidant and immunomodulatory effects of phenolic extracts from both pale (P1 - P4) and black (B1 – B4) BSG were examined. Black BSG extracts significantly (P < 0.05) protected against DNA damage induced by hydrogen peroxide (H2O2) and extracts with the highest total phenolic content (TPC) protected against 3-morpholinosydnonimine hydrochloride (SIN-1)-induced oxidative DNA damage, measured by the comet assay. Cellular antioxidant activity assays were used to measured antioxidant potential in the U937 cell line. Extracts P1 – P3 and B2 - B4 demonstrated significant (P < 0.05) antioxidant activity, measured by the superoxide dismutase (SOD) activity, catalase (CAT) activity and gluatathione (GSH) content assays. Phenolic extracts P2 and P3 from pale BSG possess anti-inflammatory activity measured in concanavalin-A (conA) stimulated Jurkat T cells by an enzyme-linked immunosorbent assay (ELISA); significantly (P < 0.05) reducing production of interleukin-2 (IL-2), interleukin-4 (IL-4, P2 only), interleukin-10 (IL-10) and interferon-γ (IFN-γ). Black BSG phenolic extracts did not exhibit anti-inflammatory effects in vitro. Hydroxycinnamic acids (HA) have previously been shown to be the phenolic acids present at highest concentration in BSG; therefore the HA profile of the phenolic extracts used in this research, the original barley (before brewing) and whole BSG was characterised and quantified using high performance liquid chromatography (HPLC). The concentration of HA present in the samples was in the order of ferulic acid (FA) > p-coumaric acid (p-CA) derivatives > FA derivatives > p-CA > caffeic acid (CA) > CA derivatives. Results suggested that brewing and roasting decreased the HA content. Protein hydrolysates from BSG were also screened for their antioxidant and anti-inflammatory potential. A total of 34 BSG protein samples were tested. Initial analyses of samples A – J found the protein samples did not exert DNA protective effects (except hydrolysate H) or antioxidant effects by the comet and SOD assays, respectively. Samples D, E, F and J selectively reduced IFN-γ production (P < 0.05) in Jurkat T cells, measured using enzyme linked immunosorbent assay (ELISA). Further testing of hydrolysates K – W, including fractionated hydrolysates with molecular weight < 3, < 5 and > 5 kDa, found that higher molecular weight (> 5 kDa) and unfractionated hydrolysates demonstrate greatest anti-inflammatory effects, while fractionated hydrolysates were also shown to have antioxidant activity, by the SOD activity assay. A commercially available yogurt drink (Actimel) and snack-bar and chocolate-drink formulations were fortified with the most bioactive phenolic and protein samples – P2, B2, W, W < 3 kDa, W < 5 kDa, W > 5 kDa. All fortified foods were subjected to a simulated gastrointestinal in vitro digestion procedure and bioactivity retention in the digestates was determined using the comet and ELISA assays. Yogurt fortified with B2 digestate significantly (P < 0.05) protected against H2O2-induced DNA damage in Caco-2 cells. Greatest immunomodulatory activity was demonstrated by the snack-bar formulation, significantly (P < 0.05) reducing IFN-γ production in con-A stimulated Jurkat T cells. Hydrolysate W significantly (P < 0.05) increased the IFN-γ reducing capacity of the snack-bar. Addition of fractionated hydrolysate W < 3 kDa and W < 5 kDa to yogurt also reduced IL-2 production to a greater extent than the unfortified yogurt (P < 0.05).

Micropropagation of Begonia and a study of genome stability in Begonia rex

The development of procedures and media for the micropropagation of B. rex are described. Media for the production of plantlets from a number of other Begonia hybrids are also provided. Growth analysis data is given for plants produced in vivo from leaf cuttings and in vitro from mature leaf petioles and immature leaves derived from singly and multiply recycled axenic plantlets. No significant difference was found in phenotype or quantitative vegetative characters for any of the populations assessed. The results presented from studies on the development of broad spectrum media for the propagation of a number of B. rex cultivars using axenic leaf explants on factorial combinations of hormones illustrate the major influence played by the genotype on explant response in vitro and suggest media on which a range of B. rex cultivars may be propagated. Procedures for in vitro irradiation and colchicine treatments to destabilize the B. rex genome have also been described. Variants produced from these treatments indicate the utility of in vitro procedures for the expression of induced somatic variation. Colour variants produced from irradiation treatment have been cultured and prove stable. Polyploids produced as variants from irradiation treatment have been subcultured but prove unstable. Media for the induction and proliferation of callus are outlined. The influence of callus subculture and aging on the stability of the B. rex genome is assessed by chromosomal analysis of cells, in vitro and in regenerants. The B. rex genome is destabilized in callus culture but attenuation of variation occurs on regeneration. Diploid cell lines are maintained in callus subcultures and supplementation of regenerative media with high cytokinin concentrations, casein hydrolysate or adenine failed to produce variants. Callus aging however resulted in the production of polyploids. The presence and expression of pre-existing somatic variation in B. rex pith and root tissue is assessed and polyploids have been produced from pith tissues cultured in vitro. The stability of the B. rex genome and the application of tissue culture to micropropagation and breeding of B. rex are discussed.

Deer forests, game shooting and landed estates in the South West of Ireland, 1840 - 1970

This thesis is concentrated on the historical aspects of the elitist field sports of deer stalking and game shooting, as practiced by four Irish landed ascendancy families in the south west of Ireland. Four great estates were selected for study. Two of these were, by Irish standards, very large: the Kenmare estate of over 136,000 acres in the ownership of the Roman Catholic Earls of Kenmare, and the Herbert estate of over 44,000 acres in the ownership of the Protestant Herbert family. The other two were, in relative terms, small: the Grehan estate of c.7,500 acres in the ownership of the Roman Catholic Grehan family, and the Godfrey estate of c.5,000 acres, in the ownership of the Protestant Barons Godfrey. This mixture of contrasting estate size, owner's religions, nobleman, minor aristocrat and untitled gentry should, it is argued, yield a diversity of the field sports and lifestyles of their owners, and go some way to assess the contributions, good or bad, they have bequeathed to modern Ireland. Equally, it should help in assessing what importance, if any, applied to hunting. In this context, hunting is here used in its broadest meaning, and includes deer stalking and game shooting, as well as hunting with dogs and hounds on foot and horseback. Where a specific type of hunting is involved, it is so described; for example, fox hunting, stag hunting, hare hunting. Similarly, the term game is sometimes used in sporting literature to encompass all species of quarry killed, and can include deer, ground game (hares and rabbits), waterfowl, and various species of game birds. Where it refers to specific species, these are so described; for example grouse, pheasants, woodcork, wild duck, etc. Since two of these estates - the Kenmare and Herbert - each created a deer forest, unique in mid-19th century Ireland, they form the core study estates; the two smaller estates serve as comparative studies. And, equally unique, as these two larger estates held the only remnant population of native Irish red deer, the survival of that herd itself forms a concomitant core area of analysis. The numerary descriptions applied to these animals in popular literature are critically reassessed against prime source historical evidence, as are the so-called deer forest 'clearances'. The core period, 1840 to 1970, is selected as the seminal period, spanning 130 years, from the creation of the deer forests to when a fundamental change in policy and administration was introduced by the state. Comparison is made with similar estates elsewhere, in Britain and especially in Scotland. Their influence on the Irish methods and style of hunting is historically examined.

Scientism and instrumentalism After the Bomb: Dr Strangelove, End Zone, Crash and The Wire

This thesis argues that through the prism of America’s Cold War, scientism has emerged as the metanarrative of the postnuclear age. The advent of the bomb brought about a new primacy for mechanical and hyperrational thinking in the corridors of power not just in terms of managing the bomb itself but diffusing this ideology throughout the culture in social sciences, economics and other such institutional systems. The human need to mitigate or ameliorate against the chaos of the universe lies at the heart of not just religious faith but in the desire for perfect control. Thus there has been a transference of power from religious faith to the apparent material power of science and technology and the terra firma these supposedly objective means supply. The Cold War, however was a highly ideologically charged opposition between the two superpowers, and the scientific methodology that sprang forth to manage the Cold War and the bomb, in the United States, was not an objective scientific system divorced from the paranoia and dogma but a system that assumed a radically fundamentalist idea of capitalism. This is apparent in the widespread diffusion of game theory throughout Western postindustrial institutions. The inquiry of the thesis thus examines the texts that engage and criticise American Cold War methodology, beginning with the nuclear moment, so to speak, and Dr Strangelove’s incisive satire of moral abdication to machine processes. Moving on chronologically, the thesis examines the diffusion of particular kinds of masculinity and sexuality in postnuclear culture in Crash and End Zone and finishing up its analysis with the ethnographic portrayal of a modern American city in The Wire. More than anything else, the thesis wishes to reveal to what extent this technocratic consciousness puts pressure on language and on binding narratives.

Corporeal prisons: dynamics of body and mise-en-scène in three films by Paul Schrader

This thesis focuses on the complex relationship between representations of the human body and the formal processes of mise-en-scène in three consecutive films by the writer-director Paul Schrader: American Gigolo (1980), Cat People (1982) and Mishima: A Life in Four Chapters (1985). While Schrader’s work has typically been critiqued under the broad category of masculinity in crisis (and often as a subset of the films of his more famous long-time collaborator, Martin Scorsese), I focus on a fiveyear early period of his filmography when he sought to explore his key themes of bodily crisis, fragmentation and alienation through an unusually intense focus upon the expressive potential of film form, specifically via the combined elements of colour, lighting, camerawork and production design. By approaching these three films as corporeal character studies of troubled figures whose emotional and psychosexual neurosis is experienced in and through the body, I will locate Schrader’s filmmaking process and style within the thematic and aesthetic contexts of both his own early film criticism and the European and Japanese art cinemas that he claims as his primary influence. In doing so, I will establish Schrader’s position as a director whose literary and theological background differentiated him from his peers of the postclassical Hollywood generation, and who thus continually sought to develop his own visual literacy through his relationship with the camera and his collaborations with more overtly style-oriented film artists. But instead of merely focusing on mise-en-scène to gain a formalist appreciation of these films, I mobilise stylistic analysis as a new critical approach towards the problematic discourses of identity and embodiment that have haunted Schrader’s career from the beginning. In particular, I argue that paying closer attention to Schrader’s formal choices sheds new light on how these films – which he approached as exercises in style – repeatedly deal with the volatile and unavoidably body-oriented categories of race, gender and sexuality. In the process, I argue that a formalist attentiveness to mise-en-scène can also provide valuable cultural insights into Schrader’s oeuvre.