Electrochemiluminescence platform for the detection of C-reactive proteins : application of recombinant antibody technology to cardiac biomarker detection

This work exploits the high-affinity of recombinant antibodies and low background electrochemiluminescence (ECL) for cardiac-biomarker detection. The developed assay is capable of fg mL-1 detection limits as well as the detection of C-Reactive Protein (CRP) over a clinically relevant range. The assay demonstrated robust reproducibility, selectivity and stability while also highlighting a novel platform for detection of cardiac biomarkers at low concentrations

Twin-tunnelling-induced ground movements in clay

Modern tunnelling methods aim to reduce ground movements arising from the construction process. In clay strata the usual method of construction is by tunnel boring machine, which allows close control of the tunnelling process; however, any movements have the potential to cause damage to existing structures at, and below, the ground surface. The construction of underground rail systems often comprises two tunnels running in opposite directions. Common practice for assessing construction-generated movements around these tunnels is to make predictions based upon individual tunnel construction and utilise superposition to generate a total deformation profile. This approach does not take into account the strain- or stress-dependent effects between tunnel constructions. A delay may result in unanticipated ground movements generated by the construction of the second tunnel. The effect of this delay on the ground movements arising between the first and the second tunnel construction process was investigated in a series of plane strain centrifuge tests. The ground movements at and below the surface were monitored and were assessed against superposition-based predictions for surface settlement with the outcomes highlighting some inconsistencies. A procedure for predicting both surface and subsurface vertical settlement profiles in the plane transverse to the advancing tunnels in clay is suggested

High Pressure Processing of Dairy Foods

End of Project ReportThe term High Pressure Processing (HPP) is used to describe the technology whereby products are exposed to very high pressures in the region of 50 - 800 MPa (500 - 8000 Atmospheres). The potential application of HPP in the food industry has gained popularity in recent years, due to developments in the construction of HPP equipment which makes the technology more affordable. Applying HPP to food products results in modifications to interactions between individual components, rates of enzymatic reactions and inactivation of micro-organisms. The first commercial HPP products appeared on the market in 1991 in Japan, where HPP is now being used commercially for products such as jams, sauces, fruit juices, rice cakes and desserts. The pioneering research into the application of HPP to milk dates back to the end of the 19th century. Application of HPP to milk has been shown to modify its gel forming characteristics as well as reducing its microbial load. HPP offers the potential to induce similar effects to those generated by heat on milk protein. Recent reports have also indicated that HPP could accelerate the ripening of cheese. Much of the Irish cheese industry is based on the production of Cheddar cheese, the ripening time for which can vary from 4 - 12 months or more, depending on grade. A substantial portion of the cost associated with Cheddar manufacture is therefore attributed to storage under controlled conditions during ripening. Thus, any technology which may accelerate the ripening of Cheddar cheese while maintaining a balanced flavour and texture is of major economic significance. While food safety is a dominant concern, consumers are increasingly demanding foods that maintain their natural appearance and flavour, while free of chemical preservatives. HPP offers the food industry the possibility of achieving these twin goals as this technology can lead to reduced microbial loads without detrimentally effecting the nutritional or sensory qualities of the product. The development of food ingredients with novel functional properties offers the dairy industry an opportunity to revitalise existing markets and develop new ones. HPP can lead to modifications in the structure of milk components, in particular protein, which may provide interesting possibilities for the development of high value nutritional and functional ingredients. Hence these projects set out to investigate the potential of HPP in the dairy industry and to identify products and processes to which it could be applied.Department of Agriculture, Food and the Marin

Release from sheep-grazing appears to put some heart back into upland vegetation:A comparison of nutritional properties of plant species in long-term grazing experiments

Rewilding or wilding is a popularised means for enhancing the conservation value of marginal land. In the British uplands, it will involve a reduction, or complete removal, of livestock grazing (sheep), based on the belief that grazing has reduced plant species diversity, the ‘Wet Desert’ hypothesis. The hope is that if livestock is removed, diversity will recover. If true, we hypothesise that the species extirpated/reduced by grazing and then recover on its removal would more nutritious compared to those that persisted. We test this hypothesis at Moor House National Nature Reserve (North‐Pennines), where seven sets of paired plots were established between 1953 and 1967 to compare ungrazed/sheep‐grazed vegetation. Within these plot‐pairs, we compared leaf properties of seven focal species that occurred only, or were present in much greater abundance, in the absence of grazing to those of 10 common species that were common in both grazed and ungrazed vegetation. Each sample was analysed for macro‐nutrients, micro‐nutrients, digestibility, palatability and decomposability. We ranked the species with respect to 22 variables based on effect size derived from Generalised Linear Modelling (GLM) and compared species using a Principal Components Analysis. We also assessed changes in abundance of the focal species through time using GLMs. Our results support the ‘Wet Desert’ hypothesis, that is, that long‐term sheep grazing has selectively removed/reduced species like our focal ones and on recovery, they were more nutritious (macro‐nutrients, some micro‐nutrients) palatable, digestible and decomposable than common species. Measured changes in abundance of the focal species suggest that their recovery will take 10–20 years in blanket bog and 60 years in high‐altitude grasslands. Collectively, these results suggest that sheep grazing has brought about biotic homogenization, and its removal in (re)wilding schemes will reverse this process eventually! The ‘white woolly maggots’ have eaten at least part of the heart out of the highlands/uplands, and it will take some time for recovery

Radiocarbon dates from jar and coffin burials of the Cardamom Mountains reveal a unique mortuary ritual in Cambodia's late- to post-Angkor period (15th-17th centuries AD)

We present the first radiocarbon dates from previously unrecorded, secondary burials in the Cardamom Mountains, Cambodia. The mortuary ritual incorporates nautical tradeware ceramic jars and log coffins fashioned from locally harvested trees as burial containers, which were set out on exposed rock ledges at 10 sites in the eastern Cardamom Massif. The suite of 28 14C ages from 4 of these sites (Khnorng Sroal, Phnom Pel, Damnak Samdech, and Khnang Tathan) provides the first estimation of the overall time depth of the practice. The most reliable calendar date ranges from the 4 sites reveals a high- land burial ritual unrelated to lowland Khmer culture that was practiced from cal AD 1395 to 1650. The time period is concurrent with the 15th century decline of Angkor as the capital of the Khmer kingdom and its demise about AD 1432, and the subsequent shift of power to new Mekong trade ports such as Phnom Penh, Udong, and Lovek. We discuss the Cardamom ritual relative to known funerary rituals of the pre to post-Angkorian periods, and to similar exposed jar and coffin burial rituals in Mainland and Island Southeast Asia

Hepatic and adipose phenotype in Alström syndrome

BACKGROUND AND AIMS: Alström syndrome (AS) is a recessive monogenic syndrome characterized by obesity, extreme insulin resistance and multi-organ fibrosis. Despite phenotypically being high risk of non-alcoholic fatty liver disease (NAFLD), there is a lack of data on the extent of fibrosis in the liver and its close links to adipose in patients with AS. Our aim was to characterize the hepatic and adipose phenotype in patients with AS. METHODS: Observational cohort study with comprehensive assessment of metabolic liver phenotype including liver elastography (Fibroscan® ), serum Enhanced Liver Fibrosis (ELF) Panel and liver histology. In addition, abdominal adipose histology and gene expression was assessed. We recruited 30 patients from the UK national AS clinic. A subset of six patients underwent adipose biopsies which was compared with control tissue from nine healthy participants. RESULTS: Patients were overweight/obese (BMI 29.3 (25.95-34.05) kg/m2 ). A total of 80% (24/30) were diabetic; 74% (20/27) had liver ultrasound scanning suggestive of NAFLD. As judged by the ELF panel, 96% (24/25) were categorized as having fibrosis and 10/21 (48%) had liver elastography consistent with advanced liver fibrosis/cirrhosis. In 7/8 selected cases, there was evidence of advanced NAFLD on liver histology. Adipose tissue histology showed marked fibrosis as well as disordered pro-inflammatory and fibrotic gene expression profiles. CONCLUSIONS: NAFLD and adipose dysfunction are common in patients with AS. The severity of liver disease in our cohort supports the need for screening of liver fibrosis in AS.Alström UKThis is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1111/liv.1316

Targeting tumour re-wiring by triple blockade of mTORC1, epidermal growth factor, and oestrogen receptor signalling pathways in endocrine-resistant breast cancer

Background Endocrine therapies are the mainstay of treatment for oestrogen receptor (ER)-positive (ER+) breast cancer (BC). However, resistance remains problematic largely due to enhanced cross-talk between ER and growth factor pathways, circumventing the need for steroid hormones. Previously, we reported the anti-proliferative effect of everolimus (RAD001-mTORC1 inhibitor) with endocrine therapy in resistance models; however, potential routes of escape from treatment via ERBB2/3 signalling were observed. We hypothesised that combined targeting of three cellular nodes (ER, ERBB, and mTORC1) may provide enhanced long-term clinical utility. Methods A panel of ER+ BC cell lines adapted to long-term oestrogen deprivation (LTED) and expressing ESR1wt or ESR1Y537S, modelling acquired resistance to an aromatase-inhibitor (AI), were treated in vitro with a combination of RAD001 and neratinib (pan-ERBB inhibitor) in the presence or absence of oestradiol (E2), tamoxifen (4-OHT), or fulvestrant (ICI182780). End points included proliferation, cell signalling, cell cycle, and effect on ER-mediated transactivation. An in-vivo model of AI resistance was treated with monotherapies and combinations to assess the efficacy in delaying tumour progression. RNA-seq analysis was performed to identify changes in global gene expression as a result of the indicated therapies. Results Here, we show RAD001 and neratinib (pan-ERBB inhibitor) caused a concentration-dependent decrease in proliferation, irrespective of the ESR1 mutation status. The combination of either agent with endocrine therapy further reduced proliferation but the maximum effect was observed with a triple combination of RAD001, neratinib, and endocrine therapy. In the absence of oestrogen, RAD001 caused a reduction in ER-mediated transcription in the majority of the cell lines, which associated with a decrease in recruitment of ER to an oestrogen-response element on the TFF1 promoter. Contrastingly, neratinib increased both ER-mediated transactivation and ER recruitment, an effect reduced by the addition of RAD001. In-vivo analysis of an LTED model showed the triple combination of RAD001, neratinib, and fulvestrant was most effective at reducing tumour volume. Gene set enrichment analysis revealed that the addition of neratinib negated the epidermal growth factor (EGF)/EGF receptor feedback loops associated with RAD001. Conclusions Our data support the combination of therapies targeting ERBB2/3 and mTORC1 signalling, together with fulvestrant, in patients who relapse on endocrine therapy and retain a functional ER

Long-term prognosis for 1-year relapse-free survivors of CD34 cell-selected allogeneic hematopoietic stem cell transplantation : a landmark analysis

Altres ajuts: This research was supported in part by National Institutes of Health award number P01 CA23766 and NIH/NCI Cancer Center Support Grant P30 CA008748. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.CD34 selection significantly improves GVHD-free survival in allogeneic hematopoietic cell transplantation (allo-HSCT). Specific information regarding long-term prognosis and risk factors for late mortality after CD34-selected allo-HSCT is lacking, however. We conducted a single-center landmark analysis in 276 patients alive without relapse 1 year after CD34-selected allo-HSCT for AML (n=164), ALL (n=33), or MDS (n=79). At 5 years' follow-up after the 1-year landmark (range 0.03-13 years), estimated RFS was 73% and OS 76%. The 5-year cumulative incidence of relapse and NRM were 11% and 16%, respectively. In multivariate analysis, HCT-CI score ≥ 3 correlated with marginally worse RFS (HR 1.78, 95% CI 0.97-3.28, p=0.06) and significantly worse OS (HR 2.53, 95% CI 1.26-5.08, p=0.004). Despite only 24% of patients with acute GVHD within 1 year, this also significantly correlated with worse RFS and OS, with increasing grades of acute GVHD associating with increasingly poorer survival on multivariate analysis (p<0.0001). Of 63 deaths after the landmark, GVHD accounted for 27% of deaths and was the most common cause of late mortality, followed by relapse and infection. While prognosis is excellent for patients alive without relapse 1 year after CD34-selected allo-HSCT, risks of late relapse and NRM persist, particularly due to GVHD

Transplantation in Remission Improves the Disease-Free Survival of Patients with Advanced Myelodysplastic Syndromes Treated with Myeloablative T Cell-Depleted Stem Cell Transplants from HLA-Identical Siblings

AbstractFrom 1985 to 2004, 49 patients with advanced myelodysplastic syndromes (MDS) (≥5% blasts) or acute myeloid leukemia (AML) transformed from MDS underwent T cell depleted bone marrow or peripheral blood hematopoietic stem cell transplantation (HSCT) from HLA-identical siblings following conditioning with a myeloablative regimen that included total body irradiation (44 patients) or busulfan (5 patients). Thirty-six patients received chemotherapy (3 low dose and 33 induction doses) before conditioning, and 13 patients did not receive any chemotherapy. Prior to transplantation, 22 of the 36 treated patients were in hematologic remission; 4 were in a second refractory cytopenia phase (26 responders); 8 had failed to achieve remission; and 2 of the responders had progression or relapse of their MDS (10 failures). No post-transplantation pharmacologic prophylaxis for graft-versus-host disease (GVHD) was given. The median age was 48 yrs (range 13-61). Forty-five of the 49 patients engrafted; 2 had primary graft failure; and 2 died before engraftment. Only 3 patients developed acute GVHD (aGVHD) (grades I and III) and 1 chronic GVHD (cGVHD). At 3 yrs post-transplantation, the overall survival (OS) was 54% in the responders; 31% in the untreated group; and 0% in the failure group (P=.0004). The disease free survival (DFS) was 50%, 15% and 0% in each group respectively (P=.0008). In multivariate analysis, disease status before cytoreduction remained highly correlated with DFS (P<.001). The cumulative incidence (CI) of relapse at 2-yrs post-transplantation for the responders was 23%; for the untreated group was 38%; and for the failures was 50%. The CI of non-relapse mortality at 2-yrs post-transplantation, for the responders was 23%; for the untreated group was 38%; and for the failures was 40%. All survivors achieved a Karnofsky Performance Status (KPS) of ≥90. These results indicate that patients with advanced MDS who achieve and remain in remission or a second refractory cytopenia phase with chemotherapy before conditioning can achieve successful long-term remissions following a myeloablative T cell depleted allogeneic HSCT