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Investigation of the substrate specificity of glutamyl endopeptidase using casein substrates
Get PDFElectrophoretically and chromatographically pure P-casein was prepared from acid caseinate exploiting the differential solubility of different caseins in the presence of Ca2+ at 4°C and 30°C. The yield of P-casein was approximately 5%. The p-casein\ud
purification process also yielded a co-fraction enriched in a-casein.GE was purified and characterized from Alcalase™ 2.4 L. Various protocols were investigated to\ud
purify GE before selecting a protocol employing hydrophobic interaction (HIC) and cation exchange (IEX) chromatography. The yield of GE obtained was approximately 42%. The purified P-casein and the enriched a-casein fractions were digested with GE at\ud
37°C and 50°C over 4 h. The results show that GE was highly specific and hydrolysed the peptide bond predominantly on the carboxy terminal of Glu residues. GE also appeared to hydrolyze bonds on the carboxy terminal o f Asp. The results indicated that Met residues were poorly preferred at the P| ’ position. Whereas, Pro residues were not preferred at the P |’ position. It was also observed that the hydrolysis of Glu-X bond in Glu-Glu-X and in Glu-Glu-Glu-X sequences (X= Arg, Asn, lie and Lys) was preferred in comparison to Glu-Glu, suggesting that at the P i’ position Glu\ud
residues were poorly preferred. Non-specific cleavages were observed at the carboxy terminal of Phe (51), Thr (128) and Gin (188) on incubation of P-casein with GE.\ud
Non-specific cleavages at the carboxy terminal of Phe (24) and Gin (131) in a sicasein were observed on incubation of the enriched a-casein with GE only at 50°C. Synthetic peptides corresponding to specific sequences of P-casein were incubated with GE at 37°C for 120 min. The LC-MS analysis on the hydrolysed sample showed that multiple phosphorylated residues near the scissile bond did not influence the substrate specificity of GE. Furthermore, these studies showed that the Glu-Pro and the Glu-Met bonds were hydrolysed
Support agencies and start-up businesses: the role of Udaras na Gaeltachta
Get PDFSmall and Medium Enterprises (SME) are the cornerstone of our economy. They\ud
provide approximately two thirds of private sector employment and contribute vastly\ud
to our exchequer. This study examines the support agencies provided to SMEs in\ud
Ireland. Investigation is primarily focused on what is important to fledgling businesses\ud
and on the ability of support agencies to meet this expectation. Possible\ud
recommendation of improvements that could potentially further enhance the assistance\ud
currently provided by support agencies was also suggested. For the purpose of my\ud
investigation Udaras Na Gaeltachta was chosen as my research subject. The results\ud
based on a face to face interview with a representative from Udaras Na Gaeltachta\ud
along with a survey of thirty of their previous clients indicates that Udaras Na\ud
Gaeltachta is responsible for providing many benefits to the businesses it assists. The\ud
initial expectations associated with the majority of the supports it provides is above\ud
seventy percent. This indicates that it has established an excellent reputation in the\ud
Gaeltacht community and is capable of providing a high quality service. However the\ud
findings also indicate that Udaras Na Gaeltachta is currently underachieving in\ud
several support areas, suggesting that to date weaknesses exist within the\ud
organisation. Further research should be considered regarding other possible\ud
recommendations for increasing the effectiveness of the support services provided by\ud
Udaras Na Gaeltachta. In addition to this, the following research topics are of interest\ud
to the researcher and if carried out would add value to this study, ‘the benefits\ud
received by the Gaeltacht community since the establishment of Udaras Na\ud
Gaeltachta’, or ‘the factors used by financial Institutes to determine which businesses\ud
qualify for funding would add value to this research’
Characterisation and investigation of alginate dressings containing hydrogen peroxide for the promotion of wound healing
Get PDFThe care of chronic wounds carries a heavy financial burden on the healthcare industry, \ud
with billons being spent annually on their treatment. This, coupled with a decreased \ud
quality of life for sufferers, has led to an urgency in developing inexpensive wound \ud
dressings that promote wound healing. Alginate gels for application as wound dressings \ud
were formed by varying alginate (0-6 %w/v), calcium carbonate (0-1 %w/v), hydrogen \ud
peroxide (0-3.75 % v/v) and hyaluronic acid (0-1.25 mg/L) content. The aging effects on \ud
the physical properties of the gels over a 14 day period were also investigated. The \ud
results indicated that the concentration of calcium carbonate and hydrogen peroxide, as \ud
well as sample age, all had a significant effect on the rupture characteristics and gelation \ud
time of the gels. Increased calcium carbonate content caused an increase in rupture force \ud
values, whereas increased hydrogen peroxide content and sample age resulted in a \ud
decrease in rupture force measurements. Increased calcium carbonate and hydrogen \ud
peroxide content produced a decrease in the time required for gel formation. Statistical \ud
models were also produced to provide a means of estimating rupture characteristics and \ud
gelation times for gels containing other concentrations of these components. Chronic \ud
wounds endure a state of hypoxia that impairs the healing process. Thus, delivery of \ud
oxygen to the wound may benefit the healing of wounds. Gels containing hydrogen \ud
peroxide were shown to release oxygen when in contact with an artificial wound \ud
containing catalase. Gels not containing hydrogen peroxide did not release oxygen when \ud
in contact with the artificial wound. The incorporation of hyaluronic acid into the gel had \ud
no significant effect of the rate of oxygen release. These gels serve as an attractive \ud
vehicle for the delivery of oxygen to wounds, thus aiding in the healing process. \ud
Culturing synovial fibroblasts in the presence of the alginate gels had a detrimental effect \ud
on the cell migration and proliferation of these cells. Cell migration and proliferation are \ud
important phases in wound healing. These findings appear to suggest that the alginate \ud
gels are not beneficial in the wound healing. However, it is more likely that these results \ud
are due to the delicate nature of the in vitro system used for the assessment of the gels
The interaction between A20 and the NR4A subfamily of nuclear receptors in the pathogenesis of rheumatoid arthritis
Get PDFRheumatoid arthritis (RA) is a chronic, progressive inflammatory disease that affects nearly 1 % \ud
of the world’s population. RA is characterized by inflammation of the synovial joints leading to \ud
joint damage which can result in deformities and loss of function. Activation of the transcription \ud
factor NF-kB is pivotal in the pathogenesis of RA, switching on multiple proinflammatory genes. \ud
The zinc finger protein A20 inhibits induction of NF-kB activation by a variety of stimuli \ud
including the proinflammatory cytokine TNF-a in a variety of cell types, dampening the immune \ud
response. The A20-related genes ABIN-1, ABIN-2 and Cezanne have also been shown to \ud
attenuate TNF-a-induced NF-kB activation. The NR4A subfamily of orphan nuclear receptors \ud
comprising of NURR1, NUR77 and NOR-1 are ligand-independent transcription factors and \ud
evidence suggests that they may have a proinflammatory role in the RA synovium. Previous \ud
studies have demonstrated that overexpression of the NR4A subfamily members led to the \ud
induction of A20 gene expression, indicating the potential role of this subfamily in the regulation \ud
of A20 and A20-interacting proteins. This study investigated the potential interaction between \ud
A20 and the NR4A member NURR1 in the context of RA. Bioinformatic analysis of A20, \ud
ABIN-1, ABIN-2 and Cezanne identified the NR4A oinding site (NBRE site) in the promoter \ud
region of all four genes, indicating that A20 and these A20-interacting genes may be induced by \ud
the NR4A subfamily of nuclear receptors. Transient co-transfection experiments and luciferase \ud
assays were carried out on cellular models of inflammatory arthritis (fibroblast-like synoviocytes \ud
and chondrocytes). The results indicated that A20 overexpression attenuates the induction of \ud
NBRE-luciferase activity by endogenous and overexpressed NURR1 in both synoviocyte and \ud
chondrocyte cells. Results in synoviocytes revealed a dose-dependent inhibitory effect of A20 on \ud
the transcriptional activity of NURR1. Further experiments demonstrate that in synoviocytes, \ud
A20 overexpression led to suppression of NR4A driven transactivation of the interleukin (IL)-8 \ud
promoter. In chondrocytes, A20 increased NURR1 induction of the IL-8 promoter. These results \ud
indicate that A20 may play a role in modulating the transcriptional activity of NURR1. Reverse \ud
transcription (RT)-PCR analysis of synoviocytes and chondrocytes stimulated with the \ud
proinflammatory cytokine TNF-a indicated that transcripts from A20, ABIN-1 and ABIN-2 are \ud
differentially expressed, while Cezanne is not affected by TNF-a treatment in these cellular \ud
contexts. qPCR analysis of A20 and ABIN-1 mRNA expression in chondrocytes in response to \ud
TNF-a treatment confirmed the end-point PCR results obtained for this cell type
Portfolio: Is it a valid and reliable assessment instrument for academic writing skills?
Get PDFThis paper critiques one assessment strategy of a level 9 academic writing module. The module supports masters students in a School of Business by utilising a number of innovative approaches to teaching and assessing of academic writing skills. The assessment instruments include a portfolio of evidence of improvement in writing skills and a series of online discussion postings; the module is 100% continuously assessed. The use of portfolio is critiqued based on both validity and reliability of the assessment instrument, and also on conformity with the concept of formative assessment
Assessing acoustic and visual monitoring techniques for the conservation management of bottlenose dolphins (Tursiops truncatus) in a special area of conservation
Get PDFBottlenose dolphins are listed under Annex II of the EU Habitats Directive and thus member states must ensure they are at a “Favourable Conservation Status”, which requires ongoing population monitoring. Estimating cetacean abundance visually can be expensive and relatively difficult. Static acoustic monitoring (SAM) provides a potential alternative for non- invasive monitoring especially in adverse weather conditions and outside daylight hours. The efficacy of four sampling techniques used to monitor bottlenose dolphins were tested in the Lower River Shannon cSAC on the west coast of Ireland for two years. Three acoustic techniques: a statically moored hydrophone and self -contained click detectors (T-PODs and C-PODs) were compared to visual surveys carried out simultaneous to acoustic monitoring, from a land-based site. “Ground-truthing“ acoustic detections with visual observations, TPOD, C-POD and hydrophone successfully detected the presence of bottlenose dolphins even though the acoustic detection rate was 1.6% (T-POD), 15.3% (C-POD) and 29.2% (hydrophone) of the visual detection rate. A significant variation was found between visual and acoustic detection rates using T-POD and hydrophone in 2008 which resulted in a higher visual detection rate compared to the T-POD (p<0.000) and hydrophone (p<0.004). The detection rate of the hydrophone was significantly higher than the detection rate of the T-POD (p<0.001). In 2009, there was also a significant difference between visual and acoustic detection rates using T-POD, C-POD and hydrophone. Again, the visual observation method recorded more detections compared to the T-POD (p<0.001), C-POD (p<0.000) and hydrophone (p<0.01). The C-POD recorded significantly less dolphin detections than the hydrophone (p<0.025) but more than the T-POD (p<0.008).\ud
Comparing the detection rates of all four methods between both study years, revealed a significant variation for the T-POD with more % of DPM/h recorded in 2009 (p<0.04). The visual and acoustic method using the hydrophone did not show any significance between 2008 and 2009 (visual: p>0.86; hydrophone: p>0.53). In 2009, the C-POD logged significantly more detections than the T-POD (p<0.008). In order to use static acoustic monitoring techniques in studies of the distribution and habitat use of cetaceans for species conservation management, it is essential to know the range over which the acoustic device effectively detects the target species in order to know the area being monitored and the number of devices to cover an area. The detection range of C-POD and statically moored hydrophone was investigated by plotting the positions of visual observations using a theodolite relative to the SAM equipment. The acoustic detection range for clicks on the C-POD was a maximum of 239 m (mean min) and 484 m (mean max), and a maximum of 639 m (mean min) and 906 m (mean max) for whistles detected by the statically moored hydrophone. \ud
The results presented are new to this area of research as no data exist on the detection distance for C-POD and bottlenose dolphins in the Shannon Estuary so far. The present study aimed to establish whether there was a relationship between group size and vocalization (whistle and “other vocalizations”) rate of bottlenose dolphins to predict group size. The objective was to examine whether the approach developed by van Parijs et al. (2002) to estimate abundances for Pacific humpback dolphins could also be applied to bottlenose dolphins. This was analyzed by investigating the correlation between acoustic activity using whistles and “other vocalizations”, and the number of bottlenose dolphins seen. The results have shown that only the whistle rate for the 25% of the most vocal active minutes correlated with group size, although even then it could still only explain 37% of the variation in whistle rate (c=0.37). Despite the constraints associated with both visual and acoustic monitoring techniques, such as false negatives (not recording or seeing animals when they are there), acoustic monitoring is the best method for nighttime surveying or for those weather conditions where visual observations are not appropriate. Automated data loggers such as the PODs can log data continuously over long periods of time and are cost-efficient due to their internal storage capacity and autonomous power supply
Steiner Waldorf education and the Irish primary curriculum : a time of opportunity
Get PDFThe object of this research was to study the implications o f Steiner Waldorf pedagogy\ud
delivered in National Schools, and to consider both its viability and usefulness. This\ud
research used both qualitative and quantitative methods of primary and secondary\ud
research.\ud
A review was carried out of the literature of the Waldorf movement internationally and\ud
specific to Ireland. A history of the Waldorf movement in Ireland, as well as a brief\ud
overview of the history of Irish publicly funded education, were both created from\ud
published literature, schools' records, and websites. Interviews were conducted with all\ud
of the full time teachers at both of the temporarily recognised Steiner National Schools,\ud
Mol an Oige and Raheen Wood. Data was compiled that showed a significant drop in\ud
the Steiner Waldorf-specific background and training of newly hired teachers at the two\ud
schools over the three years since recognition, resulting today in the majority of\ud
teachers lacking any previous Steiner Waldorf training.\ud
In fulfilling this objective, it was found that the value system of Steiner schools can be a\ud
useful addition to the options created for the families of Ireland. The general aims of the\ud
Primary School Curriculum were found to be in complete accord with those of the\ud
Steiner Waldorf approach, and multiple aspects of Waldorf pedagogy were identified\ud
which can be employed to deliver the curriculum in a vibrant and creative way. Areas of\ud
conflict were found to be centred around the Steiner Waldorf Schools' ethos, which is\ud
pedagogical rather than religious, presenting a challenge to the Department of\ud
Education and Skill's need for standardised quality assurance protocols. These areas of\ud
conflict were responded to with recommendations.\ud
Ultimately, the objective of the Department should, where possible, be to support sound\ud
and creative approaches to primary education for children in Ireland. The findings of\ud
this project clearly suggest the Steiner Waldorf model is deserving of wider recognition\ud
in this regard. There remains some question, however, whether the Department of\ud
Education is prepared to grant enough latitude for Steiner National Schools to actually\ud
apply the Steiner Waldorf model. If such latitude is not granted, these schools run the\ud
risk of becoming Steiner Waldorf in name only
An investigation into the possible need for reform of the examinership process in Ireland
Get PDFSince the recession began there has been a reduced credit flow in this country. As a \ud
result many companies have found themselves insolvent. For viable companies there \ud
is an option available called examinership. This process provides a company with \ud
court protection from its creditors for a limited period. During this period the \ud
company will receive new investment and restructure. This involves the company \ud
being given the opportunity to significantly reduce its debts. In return for the writing \ud
down of the company’s debts, their creditors will receive a dividend which is funded \ud
by this new investment. This legislation was enacted in 1990 to prevent the collapse \ud
of the Goodman group. This legislation was rushed into law and amendments had to \ud
be made in 1999 to rectify problematic issues with the original legislation. However \ud
many issues still remain with this legislation. It has been described by many as too \ud
expensive and inaccessible for small companies. \ud
This thesis aims to discover whether there is a need for reform of the current \ud
legislation. It is important in recessionary times that there is an appropriate \ud
mechanism in place to help viable companies in financial difficulty to survive. This \ud
will aid in the recovery of the economy and the protection of jobs. This thesis will \ud
investigate the issues associated with the current examinership process and also how \ud
these issues may be rectified. Practitioners in this area were asked to give their \ud
opinions on the issues identified and also if they could suggest any further \ud
improvements to this process. \ud
\ud
The research found that there may not be a need for reform; however there are minor \ud
amendments that may be required. Such as a further extension to the period of court \ud
protection for large and more complex companies. The research discovered that \ud
judges consider the number of employees a company has when deciding on a \ud
company’s application for examinership even though this is not outlined in the \ud
legislation. This should not be a factor as every job is important and every job saved \ud
could save the government €14,000 per annum. Additionally the legislation makes it \ud
impossible for SME’s to enter the process due to the high costs. Holding cases in the \ud
Circuit Court would significantly reduce these costs and make the process accessible \ud
to SME’s. This research also found that the current alternatives are inadequate and \ud
can be only used in certain circumstances
