Immunity Response of Gumboro and ND Vaccination Result which Given with Time Spacing in Broiler Chicken

An experiment was planed to know  the possibility of negative effect on Gumboro vaccination program. This program has a possibility to cause “Sick” condition on fabrious bursal of broiler chicken. From that case, it need time spacing for subsequent vaccination program, for instance ND vaccination program.  Time spacing is very importance to broiler chicken for recovering that “Sick” condition because of Gumboro vaccination effect. The purpose of his research was to know the best time spacing between Gumboro and ND vaccination program on broiler chicken. An experimental  method was used in this research, and the material used was 216 broiler chickens. A 3x4 factorial arrangement (Gumboro vaccination as factor A and ND vaccination  as factor B) with three broiler chickens per unit and six replicates of each treatment combination was used in the  research. Variables tested of  this experiment HI ND antibody titer, body weight  of 4 weeks old, and the symptom appeared after vaccination program. Anava was used to analyze the data obtained and used orthogonal polynomial for subsequent analysis. The research  results showed that the best time spacing was eight days and there was no symptom  appeared after  Gumboro  vaccination program. There were no significantly influence of vaccination treatments on broiler chicken health and body weight. (Animal Production 3(2): 67-73 (2001) Key Words: Vaccination, symptom, antibody

The Effect of Turmeric (Curcuma domestica) Extract on Water Holding Capacity, Cooking Loss, pH Values and Tenderness of Broiler Chicken Meat

Abstract.  This study was aimed to determine the influences of turmeric extract supplementation on water holding capacity, cooking loss, pH value and tenderness of broiler chicken meat Data analysis was subject to completely randomized design 5 treatments namely T0, T1, T2, T3 and T4 containing non-turmeric extract, 100 mg/kgBW/day, 200 mg/kgBW/day, 300 mg/kgBW/day and 400 mg/kgBW/day, respectively. Each unit of experiment administered 3 heads with four replications. The results indicated no effect from turmeric extract supplementation on water holding capacity, cooking loss, pH value and tenderness of broiler chicken meat. The average treatments of T0, T1, T2, T3, T4 had water holding capacities of  39.86, 37.58, 36.41, 36.94, respectively; cooking losses of 26.00, 27.58, 27.57, 27.11, and 27.49%, respectively; tenderness of 1.97, 1.95, 1.63, 1.77 and 1.99 Nmm, respectively, and final Body weights of 1,618.5, 1,568, 1,692.5, 1,651.75 and 1,462 g/head, respectively. However, a highly significant influence was observed on the pH values of 6.46, 6.04, 6.21, 6.08 and 5.98. The results indicated that none of the turmeric extract supplementation increased water holding capacity, cooking losses, tenderness and body weight. Key words: broiler, cooking loss, pH values, tenderness, water holding capacity, turmeric Abstrak. Penelitian ini bertujuan untuk mengetahui pengaruh pemberian ekstrak kunyit terhadap daya ikat air, susut masak, nilai pH dan keempukan daging ayam broiler. Manfaat penelitian yaitu tersedianya informasi ilmiah tentang ekstrak kunyit terhadap danging ayam broiler. Perlakuan yang diterapkan adalah T0 tanpa ekstrak kunyit, T1 100, T2 200, T3 300, dan T4 400 mg/kgBB/hari. Data analisis yang digunakan adalah rancangan acak lengkap yang terdiri dari 5 perlakuan dan 4 replikasi. Setiap unit percobaan terdiri dari 3 ekor. Hasil penelitian menunjukkan tidak ada pengaruh pemberian ekstrak kunyit terhadap daya ikat air, susut masak, dan keempukan daging ayam broiler. Rataan untuk perlakuan T0, T1, T2, T3, T4 pada Daya Ikat Air masing–masing 39,86; 37,58; 36,41; 36,94; 34,78%; susut masak 26,00; 27,58; 27,57; 27,11; 27,49%, keempukan 1,97; 1,95; 1,63; 1,77; 1,99 Nmm, dan bobot badan akhir 1.618,5; 1.568; 1.692,5; 1.651,75; 1.462g/ekor. Namun, memberikan pengaruh sangat nyata pada nilai pH 6,46; 6,04; 6,21; 6,08; 5,98. Hasil  menunjukkan bahwa pemberian ekstrak kunyit  tidak meningkatkan daya ikat air, susut masak, keempukan dan bobot badan. Kata Kunci : Broiler, Susut Masak, pH, Keempukan, Daya Ikat Air, Kunyit

The Growth of Muscle Cell of Inbred Chicken and Indigeneous Chicken Embryo in The Medium of Rabbit Serum and Sheep Serum

An experiment on the growth embryonic muscle cell in the rabbit and sheep serum media was conducted in the Biotechnology Laboratory of Gadjah Mada University, Yogyakarta. The aim of this experiment was to observe the potency of the growth of embryonic muscle cell of the inbred chicken and indigeneous chicken in the medium of rabbit and sheep serum. Two kinds of embryo, the inbred and indigeneous chicken of eleven days old were used in the experiment. The rabbit and the sheep serum were prepared in the laboratory. The experiment was conducted by applying Nested Classification with basic Complete Randomized Design (CRD). Data collected was analyzed using analysis of variance and also using a proliferation index formula. Samples used in those research were the inner and outer cell nucleus after fourty eight hours of the growth. The result of the experiment indicated that the index of proliferation of embryonic muscle cell of the inbred chicken in the rabbit and sheep serum were 89.65 and 84.92 percent respectively. Whereas, the proliferation index of embryonic muscle cell of the indigeneous chicken in the rabbit and sheep serum were 86.20 and 84.82 percent respectively. The total of inner muscle cell nuclei of inbred chicken embryos was significantly higher (P<0.01) than those of indigeneous chicken embryos either in the rabbit or sheep serum, but there was no difference between the serum (P>0.05). inconclusion the muscle cell of inbred and indigeneous chicken embryos could growth in both serum but the growth muscle cell of inbred chicken embryo was better than that of indigeneous chicken embryo. (Animal Production 2(2): 75-82 (2000) Key words : tissue culture, chicken embryos, index proliferation, serum.

Analysis of Local Chicken Entreprise in DAS Serayu Banyumas

The Objectives of this research was to know income and efficiency level of local chicken entreprise. Beside that, to know potency of local chicken enterprise developing in DAS Serayu, Banyumas and know factors can effect level of that income and efficiency. Methode that used at this research is survey method to farmer families. Take of research data by random sampling.The data is analysed by multiple regression analysis. The results of this research showed that income level of local chicken entreprise at DAS Serayu is Rp 277.375,00 / year and economi efficiency 2.80 , that means the farmers get return Rp 2.80 for every one unit cost addition. The age of farmers and total of chicken possession effect at efficiency of  local chicken entreprise. Potency of local chicken developing very big if showed from power of area and human resources. Very important to increase entreprise capital and increase knowledge for farmer. Beside that more important present motivation and support for develop there enterprise (Animal Production 2(1): 13-17 (2000)Key Words: local chicken, farmers income, economic efficiency

The Effect of Level Testosteron e Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm

The research entitled “The Effect of Level Testosterone Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm” was conducted in Laboratory of Physiology and Reproduction, Faculty of Animal Science, UNSOED, started at August 15th to September 15th , 2001. The aims of  this research were to obtain influences of level testosterone and dilution on speed movement and secondary abnormality kedu chicken sperm and obtain interaction between the treatments. The tapped sperm from nine kedu chickens were used in this research. This Experiment was performed 4 x 3 Factorials with Randomized  Completely Block Design (RCBD) as the basic design. The treatment combinations were level testosterone 0, 300, 600 and 900 μg (t0, t1, t2 and t3) and level of dilution 4, 6 and 8 time (in a row p1, p2 and p3). The tapping period was replicated four times as a group (replicated) with two days interval. The result of this research showed that the interaction between level of testosterone addition and level of dilution gave a non-significant effect to speed movement sperm but significant to abnormality of kedu chicken sperm. The group gave a significant influence (P<0.05) to speed movement sperm and non significant to sperm abnormality. The interaction of level testosterone addition and level dilution of kedu chicken semen (T x P) has a quadratic regression to sperm abnormality with regression comparison is Y = 24.418 – 0.014 X +1.187E – 05 X², with peak point is (543.76: 20.23) of correlation coefficient (r) 0.55 and determination coefficient (R²) as 30.34%. The addition of 600 μg testosterone level with 6 level dilutions was the best to defend sperm abnormality. (Animal Production 4(2): 60-70 (2002) Key Words : Spermatozoa, Kedu Chicken, Testosterone, Dilution, Speed Movement and Abnormality

Quality of Chicken Feather Processed in Different Conditions

Abstract. The objective of this research was to evaluate the hydrolyzed chicken feather based on pepsin digestibility and nutrient content, after physico-chemical and biological process. It was carried out by experimental methods at feed and nutrition laboratory. The treatments were hydrolyzed feather meals immersed in 0.5% NaOH and Na2S solution for 0, 2, 4, 6 and 8 hours, each treatment was repeated three times. The results showed that chemical treatment (NaOH-Na2S) in various time of incubation at 60oC followed by fermentation using Bacillus sp. MTS at 37oC for four days decreased the protein of hydrolyzed feather (78.88 to 73.06%), but increased the keratin fiber (1.9 to 3.26%). Pepsin digestibility informed that the increasing incubation time from 0, 2, 4, 6 to 8 hours resulted in higher solubility than that of control (30.2% at 8 hours vs 15.4% at 0 hours). Processing chicken feather  by  0.5% NaOH and Na2S solution at 60oC for 6 hours followed by fermentation increased the value of pepsin digestibility.  Key words: hydrolyzed, Bacillus sp. MTS, feather, solubility Abstrak. Penelitian ini bertujuan mengevaluasi kualitas nutrien tepung bulu ayam hasil proses hidrolisis secara fisiko-kimia dan biologis menggunakan Bacillus sp. MTS. Metode eksperimental digunakan dalam penelitian yang menggunakan dua tahap proses hidrolisis yaitu tahap 1: setelah perebusan bulu dalam larutan NaOH maka bulu direndam dalam larutan  0.5% NaOH dan Na2S pada 600C dan tahap 2: fermentasi bulu selama empat hari pada suhu 370C. Perlakuan berupa waktu inkubasi yaitu 0, 2, 4, 6 dan 8 jam diterapkan pada tahap kedua dengan ulangan sebanyak tiga kali. Perlakuan fisiko-kimia yang dilanjutkan fermentasi menggunakan bakteri spesifik penghasil enzim-enzim pendegradasi keratin bulu menurunkan kadar  protein tepung bulu  (78,88% menjadi 73,06%) dan meningkatkan kadar serat tepung bulu (1,9 menjadi 3,26%). Uji kelarutan protein tepung bulu dalam pepsin menginfromasikan bahwa proses tahap 1 menghasilkan nilai kelarutan protein tepung bulu yang meningkat dua kali dibanding kontrol (30,2% pada 8 jam vs 15,4% pada 0 jam inkubasi) atau enam kali dibanding tepung bulu tanpa hidrolisis (5%). Pengolahan bulu ayam menggunakan cara pemanasan, perendaman dalam larutan NaOH dan Na2S selama 6 jam pada 600C serta fermentasi menghasilkan tepung bulu dengan daya larut dalam pepsin  lebih baik dibanding tanpa pengolahan.  Kata kunci: hidrolisis, tepung-bulu, Bacillus sp. MTS, kelarutan

Fat and Cholesterol Content of Breast and Thigh Meat of Local Fowls

The research was carried out to evaluate the influence of breed on fat and cholesterol content of breast and thigh meat of local fowls.  Tree breeds of local fowl consist of six male kampong chickens, Tegal ducks and muscovy ducks were applied.  An experimental method was used in this research.  Nested design was used.  The breeds of local fowl were as treatment, the part of carcass was as sub treatment and sample was as replication.  The result of the experiment showed that the part of carcass (breast and thigh) influenced meat fat and cholesterol content.  Breast meat had higher fat and cholesterol content than thigh meat.  Meat fat content had positive correlation with meat cholesterol. (Animal Production 5(2): 79-82 (2003) Key Words: Kampung Chicken, Duck, Muscovy, Breast, Thigh, Meat Fat and Meat Cholesterol

Carcass Evaluation of Crossbred Betweeen Cockerel Kampung Chicken and Layer Hen

The research was conducted to the characteristics of carcass evaluate crossbred between cockerel of kampung chicken and Lohman layer hen. Sixteen crossbred chickens and 16 kampung chickens were reared under a similar management from 2 - 12 weeks old. The chickens were given a commercial feed which contains 21% crude protein at 2 - 4 week old, and 14% crude protein and metabolizable energy 2800 kcal/kg at 4 - 12 weeks old. The data of carcass weight, rear back weight, fore back weight, breast weight, and thigh weight were collected. Meat and bone on breast and thigh were separated. The results showed that with under similar management and feeding, the carcass values of crossbred chicken were not significantly different with kampung chicken. The crossbred chickens have meat production rate of 2.83 times as compared to kampung chicken. Higher crude protein than 14% with a balance metabolizable energy will increase the quantity and quality of crossbred chicken carcass. (Animal Production 4(2): 71-76 (2002) Key Words : Carcass, Crossbred Kampung Chicken

The Effects of Various Types of Feed additives on The Chemical Composition of Local Chicken Meat (Gallus domestica).

Abstract. This study aims to determine the effect of giving various types of feed additives to the chemical composition value of super chicken chicken (Gallus domestica). This research is an experimental research using 20 super chicken chickens that come from chicken growth research (growth study). The design used in this study was Completely Randomized Design (RAL), consisting of 4 treatments and 5 replications. The treatment given was (A0 = control (Vita chick 0.7 gram / liter; A1 = 20 ml / liter probio-FM; A2 = 0.08% MOS-oligosaccharide / kg of feed and A3 = herbal leuser KI 5 ml / liter). is a 90 day old super chicken breast Chicken Variable observed moisture content, protein content and fat content The data obtained were analyzed by using vocabulary and tested further by Duncan's Multiple Range Test The results showed that treatment (P> 0,05) to the value of water content and protein of super chicken fowl.Average value of water content at each treatment A0 (69,81%), A1 (70,74%), A2 (71,56%) and A3 (71,52%) while mean value of protein A0 (18,95%), A1 (19,61%), A2 (19,01%) and A3 (19,14%)) P <0,05) to the fat content of super chicken flesh, mean of fat content were A0 (2.02%), A1 (1.49%), A2 (1.37%) and A3 (2.0%).

Supporting ICT diffusion in Australian construction organisations

Construction organisations comprise geographically dispersed virtually-linked suborganisations that work together to realise projects. They increasingly do so using information and communication technology (ICT) to communicate, coordinate their activities and to solve complex problems. One salient problem they face is how to effectively use requisite ICT tools. One important tool at their disposal is the self-help group, a body of people that organically spring up to solve shared problems. The more recognised term for this organisational form is a community of practice (COP). COPs generate knowledge networks that enhance and sustain competitive advantage and they are also used to help COP members actually use ICT tools. Etienne Wenger defines communities of practice as “groups of people informally bound together by shared expertise and passion for a joint enterprise” (Wenger and Snyder 2000, p139). This ‘chicken-or-egg’ issue about needing a COP to use the tools that are needed to effective broaden COPs (beyond co-located these groups) led us to explore how best to improve the process of ICT diffusion through construction organisations— primarily using people supported by technology that improves knowledge sharing. We present insights gained from recent PhD research results in this area. A semistructured interview approach was used to collect data from ICT strategists and users in the three large Australian construction organisations that are among the 10 or so first tier companies by annual dollar turnover in Australia. The interviewees were categorised into five organisational levels: IT strategist, implementer, project or engineering manager, site engineer and foreman. The focus of the study was on the organisation and the way that it implements ICT diffusion of a groupware ICT diffusion initiative. Several types of COP networks from the three Australian cases are identified: withinorganisation COP; institutional, implementer or technical support; project manager/engineer focussed; and collegial support. Also, there are cross-organisational COPs that organically emerge as a result of people sharing an interest or experience in something significant. Firstly, an institutional network is defined as a strategic group, interested in development of technology innovation within an organisation. This COP principally links business process domain experts with an ICT strategist.

Development of rapid and highly resolving combinatorial genotyping schemes for Campylobacter jejuni and Campylobacter coli

Campylobacter jejuni followed by Campylobacter coli contribute substantially to the economic and public health burden attributed to food-borne infections in Australia. Genotypic characterisation of isolates has provided new insights into the epidemiology and pathogenesis of C. jejuni and C. coli. However, currently available methods are not conducive to large scale epidemiological investigations that are necessary to elucidate the global epidemiology of these common food-borne pathogens. This research aims to develop high resolution C. jejuni and C. coli genotyping schemes that are convenient for high throughput applications. Real-time PCR and High Resolution Melt (HRM) analysis are fundamental to the genotyping schemes developed in this study and enable rapid, cost effective, interrogation of a range of different polymorphic sites within the Campylobacter genome. While the sources and routes of transmission of campylobacters are unclear, handling and consumption of poultry meat is frequently associated with human campylobacteriosis in Australia. Therefore, chicken derived C. jejuni and C. coli isolates were used to develop and verify the methods described in this study. The first aim of this study describes the application of MLST-SNP (Multi Locus Sequence Typing Single Nucleotide Polymorphisms) + binary typing to 87 chicken C. jejuni isolates using real-time PCR analysis. These typing schemes were developed previously by our research group using isolates from campylobacteriosis patients. This present study showed that SNP + binary typing alone or in combination are effective at detecting epidemiological linkage between chicken derived Campylobacter isolates and enable data comparisons with other MLST based investigations. SNP + binary types obtained from chicken isolates in this study were compared with a previously SNP + binary and MLST typed set of human isolates. Common genotypes between the two collections of isolates were identified and ST-524 represented a clone that could be worth monitoring in the chicken meat industry. In contrast, ST-48, mainly associated with bovine hosts, was abundant in the human isolates. This genotype was, however, absent in the chicken isolates, indicating the role of non-poultry sources in causing human Campylobacter infections. This demonstrates the potential application of SNP + binary typing for epidemiological investigations and source tracing. While MLST SNPs and binary genes comprise the more stable backbone of the Campylobacter genome and are indicative of long term epidemiological linkage of the isolates, the development of a High Resolution Melt (HRM) based curve analysis method to interrogate the hypervariable Campylobacter flagellin encoding gene (flaA) is described in Aim 2 of this study. The flaA gene product appears to be an important pathogenicity determinant of campylobacters and is therefore a popular target for genotyping, especially for short term epidemiological studies such as outbreak investigations. HRM curve analysis based flaA interrogation is a single-step closed-tube method that provides portable data that can be easily shared and accessed. Critical to the development of flaA HRM was the use of flaA specific primers that did not amplify the flaB gene. HRM curve analysis flaA interrogation was successful at discriminating the 47 sequence variants identified within the 87 C. jejuni and 15 C. coli isolates and correlated to the epidemiological background of the isolates. In the combinatorial format, the resolving power of flaA was additive to that of SNP + binary typing and CRISPR (Clustered regularly spaced short Palindromic repeats) HRM and fits the PHRANA (Progressive hierarchical resolving assays using nucleic acids) approach for genotyping. The use of statistical methods to analyse the HRM data enhanced sophistication of the method. Therefore, flaA HRM is a rapid and cost effective alternative to gel- or sequence-based flaA typing schemes. Aim 3 of this study describes the development of a novel bioinformatics driven method to interrogate Campylobacter MLST gene fragments using HRM, and is called ‘SNP Nucleated Minim MLST’ or ‘Minim typing’. The method involves HRM interrogation of MLST fragments that encompass highly informative “Nucleating SNPS” to ensure high resolution. Selection of fragments potentially suited to HRM analysis was conducted in silico using i) “Minimum SNPs” and ii) the new ’HRMtype’ software packages. Species specific sets of six “Nucleating SNPs” and six HRM fragments were identified for both C. jejuni and C. coli to ensure high typeability and resolution relevant to the MLST database. ‘Minim typing’ was tested empirically by typing 15 C. jejuni and five C. coli isolates. The association of clonal complexes (CC) to each isolate by ‘Minim typing’ and SNP + binary typing were used to compare the two MLST interrogation schemes. The CCs linked with each C. jejuni isolate were consistent for both methods. Thus, ‘Minim typing’ is an efficient and cost effective method to interrogate MLST genes. However, it is not expected to be independent, or meet the resolution of, sequence based MLST gene interrogation. ‘Minim typing’ in combination with flaA HRM is envisaged to comprise a highly resolving combinatorial typing scheme developed around the HRM platform and is amenable to automation and multiplexing. The genotyping techniques described in this thesis involve the combinatorial interrogation of differentially evolving genetic markers on the unified real-time PCR and HRM platform. They provide high resolution and are simple, cost effective and ideally suited to rapid and high throughput genotyping for these common food-borne pathogens.

Environmental health risk assessment of dioxin exposure through foods in a Dioxin hot spot—Bien Hoa City, Vietnam

This study used the Australian Environmental Health Risk Assessment Framework to assess the human health risk of dioxin exposure through foods for local residents in two wards of Bien Hoa City, Vietnam. These wards are known hot-spots for dioxin and a range of stakeholders from central government to local levels were involved in this process. Publications on dioxin characteristics and toxicity were reviewed and dioxin concentrations in local soil, mud, foods, milk and blood samples were used as data for this risk assessment. A food frequency survey of 400 randomly selected households in these wards was conducted to provide data for exposure assessment. Results showed that local residents who had consumed locally cultivated foods, especially fresh water fish and bottom-feeding fish, free-ranging chicken, duck, and beef were at a very high risk, with their daily dioxin intake far exceeding the tolerable daily intake recommended by the WHO. Based on the results of this assessment, a multifaceted risk management program was developed and has been recognized as the first public health program ever to have been implemented in Vietnam to reduce the risks of dioxin exposure at dioxin hot-spots.

Entertaining the E Word : Barrie Kosky's On Ecstasy [A Review]

A review of Barrie Kosky's essay, On Ecstasy : Most of us describe the E word as a pleasant, out of this world experience—a type of boundless, artificial joy, deliberately induced by some kind of technicoloured drug. For others, it is that “lovey dovey” feeling. A spinning ceiling. Anything Lindt. For sensualist and soup connoisseur Barrie Kosky, it is easier than this. Being On Ecstasy involves, quite simply, his grandmother's chicken specialty—something warm and golden, surrendered with vegetables and a side of transcendental bliss. “A soup that took you to the beginning and end of time itself. A dazzling, pure, clear rhapsody” (7).

In vitro and in vivo examination of the cell surface glycoprotein CDCP1

A number of reports have demonstrated the importance of the CUB domaincontaining protein 1 (CDCP1) in facilitating cancer progression in animal models and the potential of this protein as a prognostic marker in several malignancies. CDCP1 facilitates metastasis formation in animal models by negatively regulating anoikis, a type of apoptosis triggered by the loss of attachment signalling from cell-cell contacts or cell-extra cellular matrix (ECM) contacts. Due to the important role CDCP1 plays in cancer progression in model systems, it is considered a potential drug target to prevent the metastatic spread of cancers. CDCP1 is a highly glycosylated 836 amino acid cell surface protein. It has structural features potentially facilitating protein-protein interactions including 14 N-glycosylation sites, three CUB-like domains, 20 cysteine residues likely to be involved in disulfide bond formation and five intracellular tyrosine residues. CDCP1 interacts with a variety of proteins including Src family kinases (SFKs) and protein kinase C ä (PKCä). Efforts to understand the mechanisms regulating these interactions have largely focussed on three CDCP1 tyrosine residues Y734, Y743 and Y762. CDCP1-Y734 is the site where SFKs phosphorylate and bind to CDCP1 and mediate subsequent phosphorylation of CDCP1-Y743 and -Y762 which leads to binding of PKCä at CDCP1-Y762. The resulting trimeric protein complex of SFK•CDCP1•PKCä has been proposed to mediate an anti-apoptotic cell phenotype in vitro, and to promote metastasis in vivo. The effect of mutation of the three tyrosines on interactions of CDCP1 with SFKs and PKCä and the consequences on cell phenotype in vitro and in vivo have not been examined. CDCP1 has a predicted molecular weight of ~90 kDa but is usually detected as a protein which migrates at ~135 kDa by Western blot analysis due to its high degree of glycosylation. A low molecular weight form of CDCP1 (LMWCDCP1) of ~70 kDa has been found in a variety of cancer cell lines. The mechanisms leading to the generation of LMW-CDCP1 in vivo are not well understood but an involvement of proteases in this process has been proposed. Serine proteases including plasmin and trypsin are able to proteolytically process CDCP1. In addition, the recombinant protease domain of the serine protease matriptase is also able to cleave the recombinant extracellular portion of CDCP1. Whether matriptase is able to proteolytically process CDCP1 on the cell surface has not been examined. Importantly, proteolytic processing of CDCP1 by trypsin leads to phosphorylation of its cell surface-retained portion which suggests that this event leads to initiation of an intracellular signalling cascade. This project aimed to further examine the biology of CDCP1 with a main of focus on exploring the roles played by CDCP1 tyrosine residues. To achieve this HeLa cells stably expressing CDCP1 or the CDCP1 tyrosine mutants Y734F, Y743F and Y762F were generated. These cell lines were used to examine: • The roles of the tyrosine residues Y734, Y743 and Y762 in mediating interactions of CDCP1 with binding proteins and to examine the effect of the stable expression on HeLa cell morphology. • The ability of the serine protease matriptase to proteolytically process cell surface CDCP1 and to examine the consequences of this event on HeLa cell phenotype and cell signalling in vitro. • The importance of these residues in processes associated with cancer progression in vitro including adhesion, proliferation and migration. • The role of these residues on metastatic phenotype in vivo and the ability of a function-blocking anti-CDCP1 antibody to inhibit metastasis in the chicken embryo chorioallantoic membrane (CAM) assay. Interestingly, biochemical experiments carried out in this study revealed that mutation of certain CDCP1 tyrosine residues impacts on interactions of this protein with binding proteins. For example, binding of SFKs as well as PKCä to CDCP1 was markedly decreased in HeLa-CDCP1-Y734F cells, and binding of PKCä was also reduced in HeLa-CDCP1-Y762F cells. In contrast, HeLa-CDCP1-Y743F cells did not display altered interactions with CDCP1 binding proteins. Importantly, observed differences in interactions of CDCP1 with binding partners impacted on basal phosphorylation of CDCP1. It was found that HeLa-CDCP1, HeLa-CDCP1-Y743F and -Y762F displayed strong basal levels of CDCP1 phosphorylation. In contrast, HeLa-CDCP1-Y734F cells did not display CDCP1 phosphorylation but exhibited constitutive phosphorylation of focal adhesion kinase (FAK) at tyrosine 861. Significantly, subsequent investigations to examine this observation suggested that CDCP1-Y734 and FAK-Y861 are competitive substrates for SFK-mediated phosphorylation. It appeared that SFK-mediated phosphorylation of CDCP1- Y734 and FAK-Y861 is an equilibrium which shifts depending on the level of CDCP1 expression in HeLa cells. This suggests that the level of CDCP1 expression may act as a regulatory mechanism allowing cells to switch from a FAK-Y861 mediated pathway to a CDCP1-Y734 mediated pathway. This is the first time that a link between SFKs, CDCP1 and FAK has been demonstrated. One of the most interesting observations from this work was that CDCP1 altered HeLa cell morphology causing an elongated and fibroblastic-like appearance. Importantly, this morphological change depended on CDCP1- Y734. In addition, it was observed that this change in cell morphology was accompanied by increased phosphorylation of SFK-Y416. This suggests that interactions of SFKs with CDCP1-Y734 increases SFK activity since SFKY416 is critical in regulating kinase activity of these proteins. The essential role of SFKs in mediating CDCP1-induced HeLa cell morphological changes was demonstrated using the SFK-selective inhibitor SU6656. This inhibitor caused reversion of HeLa-CDCP1 cell morphology to an epithelial appearance characteristic of HeLa-vector cells. Significantly, in vitro studies revealed that certain CDCP1-mediated cell phenotypes are mediated by cellular pathways dependent on CDCP1 tyrosine residues whereas others are independent of these sites. For example, CDCP1 expression caused a marked increase in HeLa cell motility that was independent of CDCP1 tyrosine residues. In contrast, CDCP1- induced decrease in HeLa cell proliferation was most prominent in HeLa- CDCP1-Y762F cells, potentially indicating a role for this site in regulating proliferation in HeLa cells. Another cellular event which was identified to require phosphorylation of a particular CDCP1 tyrosine residue is adhesion to fibronectin. It was observed that the CDCP1-mediated strong decrease in adhesion to fibronectin is mostly restored in HeLa-CDCP1-Y743F cells. This suggests a possible role for CDCP1-Y743 in causing a CDCP1-mediated decrease in adhesion. Data from in vivo experiments indicated that HeLa-CDCP1-Y734F cells are more metastic than HeLa-CDCP1 cells in vivo. This indicates that interaction of CDCP1 with SFKs and PKCä may not be required for CDCP1-mediated metastasis formation of HeLa cells in vivo. The metastatic phenotype of these cells may be caused by signalling involving FAK since HeLa-CDCP1- Y734F cells are the only CDCP1 expressing cells displaying constitutive phosphorylation of FAK-Y861. HeLa-CDCP1-Y762F cells displayed a very low metastatic ability which suggests that this CDCP1 tyrosine residue is important in mediating a pro-metastatic phenotype in HeLa cells. More detailed exploration of cellular events occurring downstream of CDCP1-Y734 and -Y762 may provide important insights into the mechanisms altering the metastatic ability of CDCP1 expressing HeLa cells. Complementing the in vivo studies, anti-CDCP1 antibodies were employed to assess whether these antibodies are able to inhibit metastasis of CDCP1 and CDCP1 tyrosine mutants expressing HeLa cells. It was found that HeLa- CDCP1-Y734F cells were the only cell line which was markedly reduced in the ability to metastasise. In contrast, the ability of HeLa-CDCP1, HeLa- CDCP1-Y743F and -Y762F cells to metastasise in vivo was not inhibited. These data suggest a possible role of interactions of CDCP1 with SFKs, occurring at CDCP1-Y734, in preventing an anti-metastatic effect of anti- CDCP1 antibodies in vivo. The proposal that SFKs may play a role in regulating anti-metastatic effects of anti-CDCP1 antibodies was supported by another experiment where differences between HeLa-CDCP1 cells and CDCP1 expressing HeLa cells (HeLa-CDCP1-S) from collaborators at the Scripps Research Institute were examined. It was found that HeLa-CDCP1-S cells express different SFKs than CDCP1 expressing HeLa cells generated for this study. This is important since HeLa-CDCP1-S cells can be inhibited in their metastatic ability using anti-CDCP1 antibodies in vivo. Importantly, these data suggest that further examinations of the roles of SFKs in facilitating anti-metastatic effects of anti-CDCP1 antibodies may give insights into how CDCP1 can be blocked to prevent metastasis in vivo. This project also explored the ability of the serine protease matriptase to proteolytically process cell surface localised CDCP1 because it is unknown whether matriptase can cleave cell surface CDCP1 as it has been reported for other proteases such as trypsin and plasmin. Furthermore, the consequences of matriptase-mediated proteolysis on cell phenotype in vitro and cell signalling were examined since recent reports suggested that proteolysis of CDCP1 leads to its phosphorylation and may initiate cell signalling and consequently alter cell phenotype. It was found that matriptase is able to proteolytically process cell surface CDCP1 at low nanomolar concentrations which suggests that cleavage of CDCP1 by matriptase may facilitate the generation of LWM-CDCP1 in vivo. To examine whether matriptase-mediated proteolysis induced cell signalling anti-phospho Erk 1/2 Western blot analysis was performed as this pathway has previously been examined to study signalling in response to proteolytic processing of cell surface proteins. It was found that matriptase-mediated proteolysis in CDCP1 expressing HeLa cells initiated intracellular signalling via Erk 1/2. Interestingly, this increase in phosphorylation of Erk 1/2 was also observed in HeLa-vector cells. This suggested that initiation of cell signalling via Erk 1/2 phosphorylation as a result of matriptase-mediated proteolysis occurs by pathways independent of CDCP1. Subsequent investigations measuring the flux of free calcium ions and by using a protease-activated receptor 2 (PAR2) agonist peptide confirmed this hypothesis. These data suggested that matriptase-mediated proteolysis results in cell signalling via a pathway induced by the activation of PAR2 rather than by CDCP1. This indicates that induction of cell signalling in HeLa cells as a consequence of matriptase-mediated proteolysis occurs via signalling pathways which do not involve phosphorylation of Erk 1/2. Consequently, it appears that future attempts should focus on the examination of cellular pathways other than Erk 1/2 to elucidate cell signalling initiated by matriptase-mediated proteolytic processing of CDCP1. The data presented in this thesis has explored in vitro and in vivo aspects of the biology of CDCP1. The observations summarised above will permit the design of future studies to more precisely determine the role of CDCP1 and its binding partners in processes relevant to cancer progression. This may contribute to further defining CDCP1 as a target for cancer treatment.

Socio-economic differences in takeaway food consumption among adults

To examine socioeconomic differences in the frequency and types of takeaway foods consumed. Cross-sectional postal survey. Participants were asked about their usual consumption of overall takeaway food (< four times a month, or ≥ four times a month) and 22 specific takeaway food items (< once a month, or ≥ once a month): these latter foods were grouped into “healthy” and “less healthy” choices. Socioeconomic position was measured using education and equivalised household income and differences in takeaway food consumption were assessed by calculating prevalence ratios using log binomial regression. Adults aged 25–64 years from Brisbane, Australia were randomly selected from the electoral roll (N = 903, 63.7% response rate). Compared with their more educated counterparts, the least educated were more regular consumers of overall takeaway food, fruit/vegetable juice, and less regular consumers of sushi. For the “less healthy” items, the least educated more regularly consumed potato chips, savoury pies, fried chicken, and non-diet soft drinks; however, the least educated were less likely to consume curry. Household income was not associated with overall takeaway consumption. The lowest income group were more regular consumers of fruit/vegetable juice compared with the highest income group. Among the “less healthy” items, the lowest income group were more regular consumers of fried fish, ice-cream, and milk shakes, while curry was consumed less regularly. The frequency and types of takeaway foods consumed by socioeconomically disadvantaged groups may contribute to inequalities in overweight/obesity and chronic disease.