Biomarker development for gastrointestinal and ovarian cancer: a proteomic approach

The development of new biomarkers for cancer patients would be advantageous in population screening for the early detection of cancers, pathological diagnosis, assessment of prognosis, tailoring treatment to individuals, and assessment of treatment response. With this in mind different proteomic approaches were used to identify biomarkers which could potentially aid prognosis and predict response in gastrointestinal and ovarian cancer. Raf Kinase Inhibitor Protein (RKIP) was originally purified from bovine brain extracts and named phosphatidylethanolamine-binding protein (PEBP). It has subsequently been shown to be a widely expressed and highly conserved protein. Several recent studies have suggested that RKIP may suppress metastasis in melanoma, prostate, and breast cancer, as reduction or loss of RKIP expression was observed in metastatic cell lines and metastatic tissue. In this part of the project RKIP expression was assessed by immunohistochemistry in tissue microarrays (TMA) from patients with colorectal and ovarian cancer. The results confirmed the findings of earlier studies and suggest that the level of RKIP expression is significantly and inversely associated with metastatic disease and can predict the risk of metastatic relapse in patients with no evidence of metastases at presentation. The level of RKIP expression as a prognostic factor was independent of sex, age, tumour site, mitotic index, lymphovascular invasion and tumour stage. Cytokeratin 18 (CK18) is an epithelial-specific cytokeratin that undergoes cleavage by caspases during apoptosis. Measurement of caspase-cleaved (CK18-NE) or total cytokeratin 18 (CK18) from epithelial-derived tumours could be a simple, non-invasive way to monitor or predict responses to treatment. Soluble plasma CK18-NE and CK18 were measured by ELISA from 73 patients with advanced gastrointestinal adenocarcinomas before treatment and during chemotherapy, as well as 100 healthy volunteers. Both CK18-NE and total CK18 plasma levels were significantly higher in patients compared to the healthy volunteers (p=0.015, p<0.001). The total CK18 baseline plasma levels prior to treatment were significantly higher (p=0.009) in patients who develop progressive disease than those who achieve partial response or stable disease and this correlation was confirmed in an independent validation set. The peak plasma levels of CK18 occurring in any cycle following treatment were also found to be associated with tumour response, but peak levels of CK18-NE did not reach significance (p=0.01, and p=0.07, respectively). A surface-enhanced laser desorption-ionisation mass spectrometry (SELDI-MS) pilot study on serum from 8 oesophageal cancer patients and 8 healthy volunteers revealed a novel biomarker, ~4kDa, downregulated in patients (p=0.012). An expanded 30 tumour/normal study was performed for validation which confirmed the down-regulation of this potential biomarker (p<0.0001). Attempts to identify tentatively suggested that the peptide may be inter-alpha-trypsin inhibitor heavy chain H4 precursor, which was interesting as a cleavage fragment of inter-alpha -trypsin inhibitor heavy chain H4 had been previously found to be up-regulated in patients with ovarian cancer, and down-regulated in patients with breast cancer. However, it was not possible to confidently confirm this identification. In a further part of this study, haptoglobin was found to be significantly more abundant in the serum from patients with oesophageal cancer compared to healthy volunteers. It was straightforward to isolate and identify and would be amenable to immunoassay as there are good antibodies available for confirmation. In conclusion, with the current lack of effective markers of metastatic relapse in colorectal cancer, a straightforward test like RKIP expression in the primary tumour may be a very cost-effective way to identify which patients may derive greater benefit from adjuvant treatment and closer post-operative surveillance, and in patients with advanced gastrointestinal malignancy levels of plasma CK18 are a potential marker of tumour response

Calorimetric Measurements of Magnetic-Field-Induced Inhomogeneous Superconductivity Above The Paramagnetic Limit

We report the first magneto-caloric and calorimetric observations of a magnetic-field-induced phase transition within a superconducting state to the long-sought exotic "FFLO" superconducting state first predicted over 50 years ago. Through the combination of bulk thermodynamic calorimetric and magnetocaloric measurements in the organic superconductor $\kappa$ - (BEDT-TTF)$_2$Cu(NCS)$_2$, as a function of temperature, magnetic field strength, and magnetic field orientation, we establish for the first time that this field-induced first-order phase transition at the paramagnetic limit $H_p$ for traditional superconductivity is to a higher entropy superconducting phase uniquely characteristic of the FFLO state. We also establish that this high-field superconducting state displays the bulk paramagnetic ordering of spin domains required of the FFLO state. These results rule out the alternate possibility of spin-density wave (SDW) ordering in the high field superconducting phase. The phase diagram determined from our measurements --- including the observation of a phase transition into the FFLO phase at $H_p$ --- is in good agreement with recent NMR results and our own earlier tunnel-diode magnetic penetration depth experiments, but is in disagreement with the only previous calorimetric report.Comment: 5 pages, 5 figure

CARBON BALANCE AND VEGETATION DYNAMICS IN AN OLD‐GROWTH AMAZONIAN FOREST

Amazon forests could be globally significant sinks or sources for atmospheric carbon dioxide, but carbon balance of these forests remains poorly quantified. We surveyed 19.75 ha along four 1‐km transects of well‐drained old‐growth upland forest in the Tapajós National Forest near Santarém, Pará, Brazil (2°51′ S, 54°58′ W) in order to assess carbon pool sizes, fluxes, and climatic controls on carbon balance. In 1999 there were, on average, 470 live trees per hectare with diameter at breast height (dbh) ≥10 cm. The mean (and 95% ci) aboveground live biomass was 143.7 ± 5.4 Mg C/ha, with an additional 48.0 ± 5.2 Mg C/ha of coarse woody debris (CWD). The increase of live wood biomass after two years was 1.40 ± 0.62 Mg C·ha−1·yr−1, the net result of growth (3.18 ± 0.20 Mg C·ha−1·yr−1 from mean bole increment of 0.36 cm/yr), recruitment of new trees (0.63 ± 0.09 Mg C·ha−1·yr−1, reflecting a notably high stem recruitment rate of 4.8 ± 0.9%), and mortality (−2.41 ± 0.53 Mg C·ha−1·yr−1 from stem death of 1.7% yr−1). The gain in live wood biomass was exceeded by respiration losses from CWD, resulting in an overall estimated net loss from total aboveground biomass of 1.9 ± 1.0 Mg C·ha−1·yr−1. The presence of large CWD pools, high recruitment rate, and net accumulation of small‐tree biomass, suggest that a period of high mortality preceded the initiation of this study, possibly triggered by the strong El Niño Southern Oscillation events of the 1990s. Transfer of carbon between live and dead biomass pools appears to have led to substantial increases in the pool of CWD, causing the observed net carbon release. The data show that biometric studies of tropical forests neglecting CWD are unlikely to accurately determine carbon balance. Furthermore, the hypothesized sequestration flux from CO2 fertilization (\u3c0.5 Mg C·ha−1·yr−1) would be comparatively small and masked for considerable periods by climate‐driven shifts in forest structure and associated carbon balance in tropical forests

Thrombocytosis and abnormal liver enzymes:A trigger for investigation of underlying malignancy

BackgroundThrombocytosis is often an incidental finding in primary care with a range of causes. Despite evidence of a strong association between thrombocytosis and malignancy, guidelines for investigating thrombocytosis in the absence of red flag symptoms remain unclear. A novel automated system of laboratory analysis, intelligent Liver Function Testing (iLFT), launched in Tayside in 2018 and has identified a patient group with thrombocytosis and abnormal liver test (LFT) results. This study analysed the outcome of these patients and investigated the use of thrombocytosis combined with LFTs in predicting risk of cancer.Methods and findingsBetween August 2018 and August 2020, 6792 patients underwent iLFT, with 246 found to have both thrombocytosis and at least one abnormal LFT. A random case-matched control group of 492 iLFT patients with normal platelet count and at least one abnormal LFT was created. 7.7% (95% CI 4.7-11.8%) of patients with thrombocytosis had cancer compared to 2.0% (1.0-3.7%) of controls. Patients ConclusionsThese findings suggest a substantial increased risk of cancer in patients with thrombocytosis and raised ALP. This could be developed as an adjunct to current investigation algorithms, highlighting high-risk patients and prompting further investigation (such as computed tomography scans) where indicated

Celebrity abuse on Twitter: the impact of tweet valence, volume of abuse, and dark triad personality factors on victim blaming and perceptions of severity

Celebrities are increasingly utilizing social media platforms to establish their brand and interact with their fan base, but in doing so they often become the targets of online abuse. While such abusive acts are known to cause severe consequences in the general population little is known about how celebrity abuse is perceived by observers. This study investigated observers' impressions of the severity of online abuse on Twitter, the blame attributed to celebrities for the abuse they received, and the role of the dark triad of observers' personality factors (Machiavellianism, narcissism, and psychopathy) in these decisions. We manipulated celebrity tweet content (negative, neutral, positive) and the volume of abusive comments (high, low) the tweets received. Celebrities received more blame the more negative their initial tweet was, and incidents were perceived as least severe following a negative tweet with a high volume of abuse. Observer impressions were influenced by their dark triad personality factors. Following negative tweets, as observer narcissism increased, victim blame increased and perceived severity decreased. Following positive tweets, as observer psychopathy increased, perceived severity decreased. Results are discussed in the context of the Warranting Theory of online impression formation and the ramifications for celebrity social media use are explored

Calorimetric Measurements of Magnetic-Field-Induced Inhomogeneous Superconductivity Above The Paramagnetic Limit

We report the first magnetocaloric and calorimetric observations of a magnetic-field-induced phase transition within a superconducting state to the long-sought exotic Fulde-Ferrell-Larkin-Ovchinnikov (FFLO) superconducting state, first predicted over 50 years ago. Through the combination of bulk thermodynamic calorimetric and magnetocaloric measurements in the organic superconductor κ−(BEDT−TTF)2Cu(NCS)2 as a function of temperature, magnetic field strength, and magnetic field orientation, we establish for the first time that this field-induced first-order phase transition at the paramagnetic limit Hp is a transition to a higher-entropy superconducting phase, uniquely characteristic of the FFLO state. We also establish that this high-field superconducting state displays the bulk paramagnetic ordering of spin domains required of the FFLO state. These results rule out the alternate possibility of spin-density wave ordering in the high-field superconducting phase. The phase diagram determined from our measurements—including the observation of a phase transition into the FFLO phase at Hp—is in good agreement with recent NMR results and our own earlier tunnel-diode magnetic penetration depth experiments but is in disagreement with the only previous calorimetric report

Concurrent and long-term associations between the endometrial microbiota and endometrial transcriptome in postpartum dairy cows

peer-reviewedBackground Fertility in dairy cows depends on ovarian cyclicity and on uterine involution. Ovarian cyclicity and uterine involution are delayed when there is uterine dysbiosis (overgrowth of pathogenic bacteria). Fertility in dairy cows may involve a mechanism through which the uterine microbiota affects ovarian cyclicity as well as the transcriptome of the endometrium within the involuting uterus. The hypothesis was that the transcriptome of the endometrium in postpartum cows would be associated with the cyclicity status of the cow as well as the microbiota during uterine involution. The endometrium of first lactation dairy cows was sampled at 1, 5, and 9 weeks postpartum. All cows were allowed to return to cyclicity without intervention until week 5 and treated with an ovulation synchronization protocol so that sampling at week 9 was on day 13 of the estrous cycle. The endometrial microbiota was measured by 16S rRNA gene sequencing and principal component analysis. The endometrial transcriptome was measured by mRNA sequencing, differential gene expression analysis, and Ingenuity Pathway Analysis. Results The endometrial microbiota changed from week 1 to week 5 but the week 5 and week 9 microbiota were similar. The endometrial transcriptome differed for cows that were either cycling or not cycling at week 5 and cyclicity status depended in part on the endometrial microbiota. Compared with cows cycling at week 5, there were large changes in the transcriptome of cows that progressed from non-cycling at week 5 to cycling at week 9. There was evidence for concurrent and longer-term associations between the endometrial microbiota and transcriptome. The week 1 endometrial microbiota had the greatest effect on the subsequent endometrial transcriptome and this effect was greatest at week 5 and diminished by week 9. Conclusions The cumulative response of the endometrial transcriptome to the microbiota represented the combination of past microbial exposure and current microbial exposure. The endometrial transcriptome in postpartum cows, therefore, depended on the immediate and longer-term effects of the uterine microbiota that acted directly on the uterus. There may also be an indirect mechanism through which the microbiome affects the transcriptome through the restoration of ovarian cyclicity postpartum