Hairless Streaks in Cattle Implicate TSR2 in Early Hair Follicle Formation

Four related cows showed hairless streaks on various parts of the body with no correlation to the pigmentation pattern. The stripes occurred in a consistent pattern resembling the lines of Blaschko. The non-syndromic hairlessness phenotype observed occurred across three generations of a single family and was compatible with an X-linked mode of inheritance. Linkage analysis and subsequent whole genome sequencing of one affected female identified two perfectly associated non-synonymous sequence variants in the critical interval on bovine chromosome X. Both variants occurred in complete linkage disequilibrium and were absent in more than 3900 controls. An ERCC6L missense mutation was predicted to cause an amino acid substitution of a non-conserved residue. Analysis in mice showed no specific Ercc6l expression pattern related to hair follicle development and therefore ERCC6L was not considered as causative gene. A point mutation at the 5'-splice junction of exon 5 of the TSR2, 20S rRNA accumulation, homolog (S. cerevisiae), gene led to the production of two mutant transcripts, both of which contain a frameshift and generate a premature stop codon predicted to truncate approximately 25% of the protein. Interestingly, in addition to the presence of both physiological TSR2 transcripts, the two mutant transcripts were predominantly detected in the hairless skin of the affected cows. Immunohistochemistry, using an antibody against the N-terminal part of the bovine protein demonstrated the specific expression of the TSR2 protein in the skin and the hair of the affected and the control cows as well as in bovine fetal skin and hair. The RNA hybridization in situ showed that Tsr2 was expressed in pre- and post-natal phases of hair follicle development in mice. Mammalian TSR2 proteins are highly conserved and are known to be broadly expressed, but their precise in vivo functions are poorly understood. Thus, by dissecting a naturally occurring mutation in a domestic animal species, we identified TSR2 as a regulator of hair follicle development.

Delivery of the p67 sporozoite antigen of Theileria parva by using recombinant Salmonella dublin: secretion of the product enhances specific antibody responses in cattle

The p67 sporozoite antigen of Theileria parva has been fused to the C-terminal secretion signal of Escherichia coli hemolysin and expressed in secreted form by attenuated Salmonella dublin aroA strain SL5631. The recombinant p67 antigen was detected in the supernatant of transformed bacterial cultures. Immunization trials in cattle revealed that SL5631 secreting the antigen provoked a 10-fold-higher antibody response to p67 than recombinant SL5631 expressing but not secreting p67. Immunized calves were challenged with a 80% lethal dose of T. parva sporozoites and monitored for the development of infection. Two of three calves immunized intramuscularly with the p67-secreting SL5631 strain were found to be protected, whereas only one of three animals immunized with the nonsecreting p67-expressing SL5631 strain was protected. This is the first demonstration that complete eukaryotic antigens fused to the C-terminal portion of E. coli hemolysin can be exported from attenuated Salmonella strains and that such exported antigens can protect cattle against subsequent parasite challenge.

The Cell Agglutination Agent, Phytohemagglutinin-L, Improves the Efficiency of Somatic Nuclear Transfer Cloning in Cattle (Bos taurus)

One of the several factors that contribute to the low efficiency of mammalian somatic cloning is poor fusion between the small somatic donor cell and the large recipient oocyte. This study was designed to test phytohemagglutinin (PHA) agglutination activity on fusion rate, and subsequent developmental potential of cloned bovine embryos. The toxicity of PHA was established by examining its effects on the development of parthenogenetic bovine oocytes treated with different doses (Experiment 1), and for different durations (Experiment 2). The effective dose and duration of PHA treatment (150 microg/mL, 20 min incubation) was selected and used to compare membrane fusion efficiency and embryo development following somatic cell nuclear transfer (Experiment 3). Cloning with somatic donor fibroblasts versus cumulus cells was also compared, both with and without PHA treatment (150 microg/mL, 20 min). Fusion rate of nuclear donor fibroblasts, after phytohemagglutinin treatment, was increased from 33 to 61% (P < 0.05), and from 59 to 88% (P < 0.05) with cumulus cell nuclear donors. The nuclear transfer (NT) efficiency per oocyte used was improved following PHA treatment, for both fibroblast (13% versus 22%) as well as cumulus cells (17% versus 34%; P < 0.05). The cloned embryos, both with and without PHA treatment, were subjected to vitrification and embryo transfer testing, and resulted in similar survival (approximately 90% hatching) and pregnancy rates (17-25%). Three calves were born following vitrification and embryo transfer of these embryos; two from the PHA-treated group, and one from non-PHA control group. We concluded that PHA treatment significantly improved the fusion efficiency of somatic NT in cattle, and therefore, increased the development of cloned blastocysts. Furthermore, within a determined range of dose and duration, PHA had no detrimental effect on embryo survival post-vitrification, nor on pregnancy or calving rates following embryo transfer.

Epidemiological studies of Johne’s Disease in cattle from Scottish farms, with a focus on slaughterhouse investigations

The PARABAN project has been a Scotland-wide initiative to develop and deliver farm-specific ‘best practice’ for the control of Mycobacterium avium ssp. paratuberculosis (MAP) in cattle using ‘Knowledge Exchange’. A range of partners have been involved, including nine ‘Champion Farms’. With input from the farmer, his/her vet and PARABAN advisors, a tailored monitoring and control programme was devised for each ‘Champion Farm’, taking into account the history of the disease on the farm, the physical facilities available and farmer objectives. Culling decisions based on live animal test results were incorporated into each farm-specific programme to complement the management programme already in place to maintain each herd. Results were analysed and discussed with all the partners throughout the project and then offered for wider scrutiny at farm open days. Feedback and questions from these open days have been used to complete the ‘Knowledge Exchange’ cycle. As a major component of the PARABAN project the author collected samples from all adult animals culled from ‘Champion Farms’ at slaughter or as fallen stock, irrespective of in-life MAP test status. These were then subjected to histopathological examination by experienced veterinary pathologists and the results compared with the results from in-life MAP testing. This was intended to evaluate the contribution slaughterhouse sampling could make towards decision making for disease control on farm and formed the main aim of this thesis. In total, samples of terminal ileum and draining lymph node were collected from three-hundred and fifty-two animals. A positive result on histopathology was defined as the presence of lesions typical of MAP and also the presence of acid-fast bacteria within the sections. There was found to be fair agreement between the overall results from histopathology and serum ELISA (Kappa = 0.33), though there appeared to be some variation in agreement between the tests on the individual ‘Champion Farms’. The presence of MAP was confirmed in seven of the eight farms which contributed animals to this study, despite sometimes prolonged efforts at controlling the disease. A separate study was undertaken to make use of the archives of the Scottish Centre for Production Animal Health and Food Safety at the Veterinary School, University of Glasgow. The archive contained records of cases from across southern Scotland and northern England. Analysis of the data generated from examination of these records suggested that MAP is widespread within the Scottish cattle herd and may well be increasing

Working memory correlates of three symptom clusters in schizophrenia

Examined whether discrete working memory deficits underlie positive, negative and disorganised symptoms of schizophrenia. 52 outpatients (mean age 37.5 yrs) with schizophrenia were studied using items drawn from the Positive and Negative Syndrome Scale (PANSS). Linear regression and correlational analyses were conducted to examine whether symptom dimension scores were related to performance on several tests of working memory function. Severity of negative symptoms correlated with reduced production of words during a verbal fluency task, impaired ability to hold letter and number sequences on-line and manipulate them simultaneously, reduced performance during a dual task, and compromised visuospatial working memory under distraction-free conditions. Severity of disorganisation symptoms correlated with impaired visuospatial working memory under conditions of distraction, failure of inhibition during a verbal fluency task, perseverative responding on a test of set-shifting ability, and impaired ability to judge the veracity of simple declarative statements. The present study provides evidence that the positive, negative and disorganised symptom dimensions of the PANSS constitute independent clusters, associated with unique patterns of working memory impairment.

Explaining usage of process modeling grammars : comparing three theoretical models in the study of two grammars

Our objective was to determine the factors that lead users to continue working with process modeling grammars after their initial adoption. We examined the explanatory power of three theoretical models of IT usage by applying them to two popular process modeling grammars. We found that a hybrid model of technology acceptance and expectation-confirmation best explained user intentions to continue using the grammars. We examined differences in the model results, and used them to provide three contributions. First, the study confirmed the applicability of IT usage models to the domain of process modeling. Second, we discovered that differences in continued usage intentions depended on the grammar type instead of the user characteristics. Third, we suggest implications and practice.

Contexts, hybrids and network governance : a comparison of three case-studies in infrastructure governance

While hybrid governance arrangements have been a major element of organisational architecture for some time, the contemporary operating environment has brought to the fore new conditions and expectations for the governance of entities that span conventional public sector departments, private firms and community organisations or groups. These conditions have resulted in a broader array of mixed governance configurations including Public Private Partnerships, alliances, and formal and informal collaborations. In some such arrangements, market based or ‘complete’ contractual relationships have been introduced to replace or supplement existing traditional ‘hierarchical’ and/or newer relational ‘network-oriented’ institutional associations. While there has been a greater reliance on collaborative or relational contracts as an underpinning institutional model, other modes of hierarchy and market may remain in operation. The success of these emergent hybrid forms has been mixed. There are examples of hybrids that have been well adopted, achieving the desired goals of efficiency, effectiveness and financial accountability; while others have experienced implementation problems which have undermined their results. This paper postulates that the cultural and institutional context within which hybrids operate may contribute to the implementation processes employed and the level of success attained. The paper explores hybrid arrangements through three cases of the use of inter-organisational arrangements in three different national contexts. Distilling the various elements of hybrids and the impact of institutional context will provide important insights for those charged with the responsibility for the formation and key infrastructure and public value development.

Physical activity in three regional communities in Queensland

Objective. To describe physical activity participation in three Queensland regional communities. Design. Cross-sectional mail survey of randomly selected residents, stratified by age and sex. Setting. Esk, Mareeba and Mount Isa. Participants. 1219 (58% female) adults, with a mean age 46.7 (SD 14.7) years. Main outcome measures. Proportion of people inactive, meeting Australian activity guidelines (a minimum of 150 minutes/week and 5 sessions/week), and walking a dog daily; time spent walking and cycling for transport; location and type of recreational physical activities. Results. Overall, 18% of respondents were inactive, with the highest proportions among women (22.3%) and older adults in Mount Isa (24.3%). The proportion meeting activity guidelines was 47% with the lowest proportions among women in Mount Isa (40.4%). Although 63% reported owning a dog, only 22% reported walking a dog daily. Few people reported walking or cycling for transport. The most common types of activities were walking, home-based exercise, running/jogging, and swimming, and the most common location was at or near home. Conclusions. Physical activity levels were lower in these regional communities than the state average. The findings indicate a need for physical activity policy and intervention strategies targeting regional and rural areas. This could focus on women and older adults, dog walking, and physical activity opportunities in or near the home.

Video-based training combined with flotation tank recovery does not improve three-point shooting in basketball

Video-based training combined with flotation tank recovery may provide an additional stimulus for improving shooting in basketball. A pre-post controlled trial was conducted to assess the effectiveness of a 3 wk intervention combining video-based training and flotation tank recovery on three-point shooting performance in elite female basketball players. Players were assigned to an experimental (n=10) and control group (n=9). A 3 wk intervention consisted of 2 x 30 min float sessions a week which included 10 min of video-based training footage, followed by a 3 wk retention phase. A total of 100 three-point shots were taken from 5 designated positions on the court at each week to assess three-point shooting performance. There was no clear difference in the mean change in the number of successful three-point shots between the groups (-3%; ±18%, mean; ±90% confidence limits). Video-based training combined with flotation recovery had little effect on three-point shooting performance.

Toxicity profile of the immunomodulatory thalidomide analogue, lenalidomide : Phase I clinical trial of three dosing schedules in patients with solid malignancies

Thalidomide is an anti-angiogenic agent currently used to treat patients with malignant cachexia or multiple myeloma. Lenalidomide (CC-5013) is an immunomodulatory thalidomide analogue licensed in the United States of America (USA) for the treatment of a subtype of myelodysplastic syndrome. This two-centre, open-label phase I study evaluated dose-limiting toxicities in 55 patients with malignant solid tumours refractory to standard chemotherapies. Lenalidomide capsules were consumed once daily for 12 weeks according to one of the following three schedules: (I) 25 mg daily for the first 7 d, the daily dose increased by 25 mg each week up to a maximum daily dose of 150 mg; (II) 25 mg daily for 21 d followed by a 7-d rest period, the 4-week cycle repeated for 3 cycles; (III) 10 mg daily continuously. Twenty-six patients completed the study period. Two patients experienced a grade 3 hypersensitivity rash. Four patients in cohort I and 4 patients in cohort II suffered grade 3 or 4 neutropaenia. In 2 patients with predisposing medical factors, grade 3 cardiac dysrhythmia was recorded. Grade 1 neurotoxicity was detected in 6 patients. One complete and two partial radiological responses were measured by computed tomography scanning; 8 patients had stable disease after 12 weeks of treatment. Fifteen patients remained on treatment as named patients; 1 with metastatic melanoma remains in clinical remission 3.5 years from trial entry. This study indicates the tolerability and potential clinical efficacy of lenalidomide in patients with advanced solid tumours who have previously received multi-modality treatment. Depending on the extent of myelosuppressive pre-treatment, dose schedules (II) or (III) are advocated for large-scale trials of long-term administration. © 2006 Elsevier Ltd. All rights reserved.