Assessing, and understanding, European organic dairy farmers' intentions to improve herd health.

Many believe the health status of organic dairy herds in Europe should be improved to meet consumers' and legislators' expectations to improve animal welfare. This paper reports on a study in four countries that examined dairy farmers' intentions towards improving the health status of their organic herds through the use of the Theory of Planned Behaviour. It was found that farmers across the countries were positive about taking additional preventative measures to improve the health status of their herds. They believed this would not only improve herd physical performance, such as milk yield and fertility, but also achieve greater cost effectiveness and improved job satisfaction for them. Most study farmers would implement a tailored package of improvement measures designed by the study team with higher uptake most likely being by younger farmers, those who make greater use of veterinarians and professional advisory services, and those supplying specialist milk-marketing chains. Furthermore, farmers will be most likely to take-up additional health promotion if compatible with their everyday activities and if they have strong business performance goals aimed at maximising the physical performance of the herd.

Using farmers' attitude and social pressures to design voluntary Bluetongue vaccination strategies.

Understanding the context and drivers of farmers' decision-making is critical to designing successful voluntary disease control interventions. This study uses a questionnaire based on the Reasoned Action Approach framework to assess the determinants of farmers' intention to participate in a hypothetical reactive vaccination scheme against Bluetongue. Results suggest that farmers' attitude and social pressures best explained intention. A mix of policy instruments can be used in a complementary way to motivate voluntary vaccination based on the finding that participation is influenced by both internal and external motivation. Next to informational and incentive-based instruments, social pressures, which stem from different type of perceived norms, can spur farmers' vaccination behaviour and serve as catalysts in voluntary vaccination schemes.

Expected utility of voluntary vaccination in the middle of an emergent Bluetongue virus serotype 8 epidemic: a decision analysis parameterized for Dutch circumstances.

In order to put a halt to the Bluetongue virus serotype 8 (BTV-8) epidemic in 2008, the European Commission promoted vaccination at a transnational level as a new measure to combat BTV-8. Most European member states opted for a mandatory vaccination campaign, whereas the Netherlands, amongst others, opted for a voluntary campaign. For the latter to be effective, the farmer's willingness to vaccinate should be high enough to reach satisfactory vaccination coverage to stop the spread of the disease. This study looked at a farmer's expected utility of vaccination, which is expected to have a positive impact on the willingness to vaccinate. Decision analysis was used to structure the vaccination decision problem into decisions, events and payoffs, and to define the relationships among these elements. Two scenarios were formulated to distinguish farmers' mindsets, based on differences in dairy heifer management. For each of the scenarios, a decision tree was run for two years to study vaccination behaviour over time. The analysis was done based on the expected utility criterion. This allows to account for the effect of a farmer's risk preference on the vaccination decision. Probabilities were estimated by experts, payoffs were based on an earlier published study. According to the results of the simulation, the farmer decided initially to vaccinate against BTV-8 as the net expected utility of vaccination was positive. Re-vaccination was uncertain due to less expected costs of a continued outbreak. A risk averse farmer in this respect is more likely to re-vaccinate. When heifers were retained for export on the farm, the net expected utility of vaccination was found to be generally larger and thus was re-vaccination more likely to happen. For future animal health programmes that rely on a voluntary approach, results show that the provision of financial incentives can be adjusted to the farmers' willingness to vaccinate over time. Important in this respect are the decision moment and the characteristics of the disease. Farmers' perceptions of the disease risk and about the efficacy of available control options cannot be neglected.

Collagen type XI α1 facilitates head and neck squamous cell cancer growth and invasion.

BACKGROUND: Although it is well established that the extracellular matrix affects tumour progression, not much is known about the various components and their effect on head and neck squamous cell carcinoma (HNSCC) progression. Levels of collagen type XI α1 (colXIα1), a minor fibrillar collagen, have been shown to be increased in tumour compared with normal tissue in several cancers, including colorectal, breast, and non-small cell lung cancer. Currently, the functional significance of colXIα1 is not understood. METHODS: We examined the expression levels of colXIα1 mRNA and elucidated the functional role of colXIα1 in HNSCC. Cell proliferation, invasion, and migration were examined with and without colXIα1 knockdown with siRNA in HNSCC cells. RESULTS: Our data demonstrate that colXIα1 expression is increased in tumour samples compared with levels in normal adjacent tissue in 16/23 HNSCC patients. In addition, colα11 is increased in HNSCC cell lines compared with normal immortalised epithelial cells and is increased in tumour-derived fibroblasts compared with normal fibroblasts. Using an siRNA approach, we demonstrate that colXIα1 contributes to proliferation, migration, and invasion of HNSCC. CONCLUSION: Our cumulative findings suggest that colXIα1 contributes to HNSCC tumorigenesis and may serve as a potential therapeutic target.

Assessment of ultraviolet-radiation-induced DNA damage within melanocytes in skin of different constitutive pigmentation.

BACKGROUND: Melanoma incidence and pigmentary disorders are known to be related to the degree of skin pigmentation, but few data exist on the specific impact of ultraviolet radiation (UVR) on melanocytes in skin of different constitutive pigmentation. OBJECTIVES: To analyse UVR-induced DNA damage within melanocytes in different skin-colour types. METHODS: Skin samples were objectively classified into light, intermediate, tan, brown and dark skin according to their individual typology angle (°ITA), based on colorimetric parameters. Samples were exposed to increasing doses of solar simulated radiation. Detection of DNA damage specifically in melanocytes was achieved by cyclobutane thymine dimer (CPD)-tyrosinase-related protein 1 double staining. RESULTS: For light, intermediate and tan skin, accumulation of CPDs in melanocytes was detected at the lowest dose, with a steep increase with dose. At estimated erythemally equivalent doses, around 80-100% of melanocytes were positive for CPDs in tan, intermediate and light skin types. In contrast, in dark and brown skin types, CPDs were found in only approximately 15% of melanocytes at the highest dose. CONCLUSIONS: This work demonstrates that melanocytes from constitutively highly pigmented skin types are less impacted in terms of UVR-induced DNA damage than those from lighter skin types, even those that are moderately pigmented.

Patient safety on the otolaryngology service: the role of an established rapid response system.

OBJECTIVE: To study the medical emergencies occurring on a tertiary otolaryngology service identified using a rapid response system (RRS). DESIGN: Retrospective chart review of RRS activations during 21 months. SETTING: Specialised otolaryngology care unit within the University of Pittsburgh Medical Center Presbyterian/Montefiore Hospital, a tertiary, academic, teaching hospital in the USA. INTERVENTION(S): None. RESULTS: 1171 unit admissions. Unit mortality was 5.1/1000 admissions. 53 patients were involved in 67 RRS activations (4/53 deaths). 32 of 67 events were due to respiratory derangements, most commonly pneumonia. 18 of 67 events were due to cardiovascular abnormalities, most commonly hypertension and myocardial infarction. 11 of 67 events were secondary to mental status changes, several of which were related to adverse drug events. 6 of 67 events were secondary to acute bleeding. 23 of 67 events occurred within 24 h of patient transfer/admission, 14 of those after operations. RRS activation was a marker for in-hospital death (RR 42.2, 95% CI 7.9 to 225.2) compared with that in patients not activating the RRS. CONCLUSIONS: Although otolaryngology care units attempt to prevent adverse events, emergencies still occur. RRSs identify deteriorating otolaryngology patients who are at increased risk for mortality. RRSs are an efficient mechanism of intervention during a medical emergency. RRSs provide a convenient method of identifying medical/system errors and educational opportunities.

Relationship between skin response to ultraviolet exposure and skin color type.

Sun exposure is responsible for detrimental damage ranging from sunburn to photoaging and skin cancer. This damage is likely to be influenced by constitutive pigmentation. The relationship between ultraviolet (UV) sensitivity and skin color type was analyzed on 42 ex vivo skin samples objectively classified from light to dark skin, based on their values of individual typology angle (ITA) determined by colorimetric parameters. The biologically efficient dose (BED) was determined for each sample by quantifying sunburn cells after exposure to increasing doses of UV solar-simulated radiation. Typical UV-induced biologic markers, other than erythema, such as DNA damage, apoptosis and p53 accumulation, were analyzed. A statistically significant correlation was found between ITA and BED and, ITA and DNA damage. Interestingly, DNA lesions were distributed throughout the whole epidermal layers and the uppermost dermal cells in light, intermediate and tanned skin while they were restricted to suprabasal epidermal layers in brown or dark skin. Our data support, at the cellular level, the relationship between UV sensitivity and skin color type. They emphasize the impact of DNA damage accumulation in basal layer in relation to the prevalence of skin cancer.

Arsenite-inducible RNA-associated protein (AIRAP) protects cells from arsenite toxicity.

Exposure of cells to arsenicals activates multiple stress pathways resulting in the induction of specific genes whose identity and role in the adaptation to arsenical-induced cellular stress are poorly understood. We report here the identification of a novel gene encoding an arsenite-inducible, cysteine- and histidine-rich RNA-associated protein, AIRAP, that is conserved among mammals, Drosophila and C elegans. Immunochemistry and cell fractionation experiments indicate that, when induced, AIRAP is present in both the nucleus and the cytoplasm, and cross-linking experiments indicate that it associates with RNA in vivo. The expression of a C elegans homologue of AIRAP, aip-1, is also induced by exposure to arsenite, and expression of an aip-1::gfp transgene is most pronounced in hypodermal cells. RNA-mediated interference (RNAi) of aip-1 lowers the resistance of nematodes to arsenite yet does not appear to affect viability under standard growth conditions. These experiments suggest a role for AIRAP/AIP-1 in protecting cells from the toxic effects of arsenite.

Portal vein thrombosis following laparoscopic splenectomy for beta-thalassemia: a case study.

Portal vein thrombosis is a rare but well-recognized complication of splenectomy. We present the case of a 31-year-old woman with transfusion-dependent b-thalassemia who underwent a laparoscopic splenectomy to reduce her transfusion requirements. Postoperatively, she developed portal vein thrombosis, diagnosed by abdominal CT scanning on postoperative day 4. After being treated with anticoagulation and antibiotic therapy, she obtained prompt resolution of her symptoms. This report summarizes the first reported incidence of portal vein thrombosis following laparoscopic splenectomy and presents the current theories regarding the etiology and treatment of postsplenectomy portal vein thrombosis.

Male sterility and enhanced radiation sensitivity in TLS(-/-) mice.

TLS (also known as FUS) is an RNA-binding protein that contributes the N-terminal half of fusion oncoproteins implicated in the development of human liposarcomas and leukemias. Here we report that male mice homozygous for an induced mutation in TLS are sterile with a marked increase in the number of unpaired and mispaired chromosomal axes in pre-meiotic spermatocytes. Nuclear extracts from TLS(-/-) testes lack an activity capable of promoting pairing between homologous DNA sequences in vitro, and TLS(-/-) mice and embryonic fibroblasts exhibit increased sensitivity to ionizing irradiation. These results are consistent with a role for TLS in homologous DNA pairing and recombination.

Human 75-kDa DNA-pairing protein is identical to the pro-oncoprotein TLS/FUS and is able to promote D-loop formation.

Homologous recombination plays a fundamental role in DNA double-strand break repair. Previously, we detected two mammalian nuclear proteins of 100 and 75 kDa (POMp100 and POMp75, respectively) that are able to promote homologous DNA pairing, a key step in homologous recombination. Here we describe the identification of human (h) POMp75 as the pro-oncoprotein TLS/FUS. hPOMp75/TLS binds both single- and double-stranded DNAs and mediates annealing of complementary DNA strands. More important, it promotes the uptake of a single-stranded oligonucleotide into a homologous superhelical DNA to form a D-loop. The formation of a D-loop is an essential step in DNA double-strand break repair through recombination. DNA annealing and D-loop formation catalyzed by hPOMp75/TLS require Mg(2+) and are ATP-independent. Interestingly, the oncogenic fusion form TLS-CHOP is not able to promote DNA pairing. These data suggest a possible role for hPOMp75/TLS in maintenance of genomic integrity.

Induction of a secreted protein by the myxoid liposarcoma oncogene.

The TLS-CHOP oncoprotein, found in the majority of human myxoid liposarcomas, consists of a fusion between the transcription factor CHOP/GADD153 and the N terminus of an RNA-binding protein TLS/FUS. Clinical correlation and in vitro transformation assays indicate that the N terminus of TLS plays an important role in oncogenesis by TLS-CHOP. Until now, however, the only activity attributed to the oncoprotein is that of inhibiting the binding of transcription factors of the C/EBP class to certain adipogenic target genes, a function that TLS-CHOP shares with the nononcogenic CHOP protein. Here we report the isolation of a gene, DOL54, that is activated in primary fibroblasts by the expression of TLS-CHOP. DOL54 is expressed in the neoplastic component of human myxoid liposarcomas and increases the tumorigenicity of cells injected in nude mice. Activation of DOL54 requires an intact DNA-binding and dimerization domain in TLS-CHOP, a suitable cellular dimerization partner, and depends on the TLS N terminus. Normal adipocytic differentiation is associated with an early and transient expression of DOL54, and the gene encodes a secreted protein that is tightly associated with the cell surface or extracellular matrix. TLS-CHOP thus leads to the unscheduled expression of a gene that is normally associated with adipocytic differentiation.

CHOP-Dependent stress-inducible expression of a novel form of carbonic anhydrase VI.

CHOP (also called GADD153) is a stress-inducible nuclear protein that dimerizes with members of the C/EBP family of transcription factors and was initially identified as an inhibitor of C/EBP binding to classic C/EBP target genes. Subsequent experiments suggested a role for CHOP-C/EBP heterodimers in positively regulating gene expression; however, direct evidence that this is the case has so far not been uncovered. Here we describe the identification of a positively regulated direct CHOP-C/EBP target gene, that encoding murine carbonic anhydrase VI (CA-VI). The stress-inducible form of the gene is expressed from an internal promoter and encodes a novel intracellular form of what is normally a secreted protein. Stress-induced expression of CA-VI is both CHOP and C/EBPbeta dependent in that it does not occur in cells deficient in either gene. A CHOP-responsive element was mapped to the inducible CA-VI promoter, and in vitro footprinting revealed binding of CHOP-C/EBP heterodimers to that site. Rescue of CA-VI expression in c/ebpbeta-/- cells by exogenous C/EBPbeta and a shorter, normally inhibitory isoform of the protein known as LIP suggests that the role of the C/EBP partner is limited to targeting the CHOP-containing heterodimer to the response element and points to a preeminent role for CHOP in CA-VI induction during stress.

Identification of novel stress-induced genes downstream of chop.

CHOP (GADD153) is a small nuclear protein that dimerizes avidly with members of the C/EBP family of transcription factors. Normally undetectable, it is expressed at high levels in cells exposed to conditions that perturb protein folding in the endoplasmic reticulum and induce an endoplasmic reticulum stress response. CHOP expression in stressed cells is linked to the development of programmed cell death and, in some instances, cellular regeneration. In this study, representational difference analysis was used to compare the complement of genes expressed in stressed wild-type mouse embryonic fibroblasts with those expressed in cells nullizygous for chop. CHOP expression, in concert with a second signal, was found to be absolutely required for the activation by stress of a set of previously undescribed genes referred to as DOCs (for downstream of CHOP). DOC4 is a mammalian ortholog of a Drosophila gene, Tenm/Odz, implicated in patterning of the early fly embryo, whereas DOC6 encodes a newly recognized homolog of the actin-binding proteins villin and gelsolin. These results reveal the existence of a novel CHOP-dependent signaling pathway, distinct from the known endoplasmic reticulum unfolded protein response, which may mediate changes in cell phenotype in response to stress.

TLS (FUS) binds RNA in vivo and engages in nucleo-cytoplasmic shuttling.

TLS, the product of a gene commonly translocated in liposarcomas (TLS), is prototypical of a newly identified class of nuclear proteins that contain a C-terminal domain with a distinct RNA recognition motif (RRM) surrounded by Arg-Gly-Gly (RGG) repeats. Its unique N terminus serves as an essential transforming domain for a number of fusion oncoproteins in human sarcomas and leukemias. In this study we use an in vivo UV crosslinking procedure to probe the interactions of TLS with RNA. TLS is found to bind RNA in vivo and the association of TLS with RNA is rapidly diminished by treating cells with transcriptional inhibitors. This suggests that the species bound by TLS turns over rapidly. Surprisingly, the RRM was found to be dispensable for RNA binding by TLS in vivo, suggesting that at any one time most of the interactions between TLS and RNA in the cell are not sequence specific. Analysis of inter specific heterokaryons formed between human and mouse or Xenopus cells revealed that TLS engages in rapid nucleocytoplasmic shuttling, a finding confirmed by the ability of anti-TLS antibodies to trap TLS when injected into the cytoplasm of HeLa cells. Cellular fractionation experiments suggest that TLS binds to RNA in both the nucleus and cytoplasm and support the hypothesis that TLS functions as a heterogeneous ribonuclear protein (hnRNP)-like chaperone of RNA. These findings are discussed in the context of the role altered forms of TLS play in cellular transformation.