Faecal contamination of lettuce heads after manure application

In recent years, an increasing number of disease outbreaks have been associated with consumption of contaminated vegetables. Thus, it has been speculated to what extent such contamination is associated with application of animal manure as fertilizer, which is particularly practiced in organic vegetable production where conventional fertilizers are prohibited. A field survey was therefore performed aiming to assess the survival and transfer of E. coli from animal manure to lettuces, with E. coli serving as an indicator of bacterial enteric pathogens. Animal manure was applied to 3 Danish fields prior to planting of lettuce seedlings, then 5-8 weeks later at the normal time of harvest, inner and outer leafs of 10 lettuce heads were pooled into one sample unit with a total of 50 pools per field. Additionally, in one field, 15 soil samples were collected weekly until the harvest time. E. coli was enumerated by plating 1 mL of 10-fold serial dilutions of 5 g of homogenized sample material, i.e. manure, soil and lettuce onto PetrifilmTM Select E. coli count plates (3M), which were then incubated 24 h at 44°C. The manure applied to the fields contained 3.0-4.5 Log10 E. coli CFU/g and E. coli was found in 36-54% of the pooled lettuce samples with a detection limit of 10 CFU/g. Numbers of E. coli in 14-20% of pooled lettuce samples exceeded a satisfactory microbiological hygiene criteria level of 100 CFU/g. The highest percentage of faecally contaminated lettuce heads (54%) coincided with the shortest growth period studied indicating that the time gap between application of manure and harvest and the survival of E. coli (and pathogens) influences the contamination of lettuce via manure amended soil. However, at the time of harvest, the numbers of E. coli in 5 of 15 soil samples were reduced below the detection limit and no samples exceeded 100 CFU/g. This is in contrast to the lettuce samples, where 20% of faecally contaminated samples had >100 E. coli/g, which may indicate that faeces contamination of crops could originate from alternative sources, such as contaminated water and wildlife. Comparisons of the genotype of isolated E. coli strains could help to elucidate this

Cyclin F Is Degraded during G2-M by Mechanisms Fundamentally Different from Other Cyclins

Cyclin F, a cyclin that can form SCF complexes and bind to cyclin B, oscillates in the cell cycle with a pattern similar to cyclin A and cyclin B. Ectopic expression of cyclin F arrests the cell cycle in G2/M. How the level of cyclin F is regulated during the cell cycle is completely obscure. Here we show that, similar to cyclin A, cyclin F is degraded when the spindle assembly checkpoint is activated and accumulates when the DNA damage checkpoint is activated. Cyclin F is a very unstable protein throughout much of the cell cycle. Unlike other cyclins, degradation of cyclin F is independent of ubiquitination and proteasome-mediated pathways. Interestingly, proteolysis of cyclin F is likely to involve metalloproteases. Rapid destruction of cyclin F does not require the N-terminal F-box motif but requires the COOH-terminal PEST sequences. The PEST region alone is sufficient to interfere with the degradation of cyclin F and confer instability when fused to cyclin A. These data show that although cyclin F is degraded at similar time as the mitotic cyclins, the underlying mechanisms are entirely distinct

Resiliencia, significado en la vida y satisfacción vital: un análisis de los efectos moderadores

While presence of meaning in life (i.e., presence) is associated with a plethora of desirable qualities (e.g., greater well-being, longevity, positive affect), search for meaning is associated with psychological distress (e.g., reports of conflict, rumination, depression; Boyle, Barnes, Buchman, & Bennett, 2009). Individuals with higher resiliency, defined as a multifaceted competency in adapting and recovering from adversity, could potentially mitigate the distress associated with search, and thus, achieve greater satisfaction with life (SWL). The present study examined the moderating role of meaning in life between resiliency (i.e., sense of mastery and sense of relatedness) and SWL in a sample of Canadian university students (N=289). Hierarchical regression analyses showed that there was a positive association between resiliency and SWL and this association was stronger at higher levels compared to lower levels of search for meaning. These results suggest that individuals searching for meaning with high levels of mastery have the greatest SWL, while their counterparts with low mastery have the lowest SWL. Similar moderating effects of search were found with the positive association between sense of relatedness and SWL. Overall, findings suggest that protective factors in resiliency may buffer against the potential negative impact of search. While presence of meaning in life (i.e., presence) is associated with a plethora of desirable qualities (e.g., greater well-being, longevity, positive affect), search for meaning is associated with psychological distress (e.g., reports of conflict, rumination, depression; Boyle, Barnes, Buchman, & Bennett, 2009). Individuals with higher resiliency, defined as a multifaceted competency in adapting and recovering from adversity, could potentially mitigate the distress associated with search, and thus, achieve greater satisfaction with life (SWL). The present study examined the moderating role of meaning in life between resiliency (i.e., sense of mastery and sense of relatedness) and SWL in a sample of Canadian university students (N=289). Hierarchical regression analyses showed that there was a positive association between resiliency and SWL and this association was stronger at higher levels compared to lower levels of search for meaning. These results suggest that individuals searching for meaning with high levels of mastery have the greatest SWL, while their counterparts with low mastery have the lowest SWL. Similar moderating effects of search were found with the positive association between sense of relatedness and SWL. Overall, findings suggest that protective factors in resiliency may buffer against the potential negative impact of search

Intravital Imaging of Adoptive T-Cell Morphology, Mobility and Trafficking Following Immune Checkpoint Inhibition in a Mouse Melanoma Model

Efficient T-cell targeting, infiltration and activation within tumors is crucial for successful adoptive T-cell therapy. Intravital microscopy is a powerful tool for the visualization of T-cell behavior within tumors, as well as spatial and temporal heterogeneity in response to immunotherapy. Here we describe an experimental approach for intravital imaging of adoptive T-cell morphology, mobility and trafficking in a skin-flap tumor model, following immune modulation with immune checkpoint inhibitors (ICIs) targeting PD-L1 and CTLA-4. A syngeneic model of ovalbumin and mCherry-expressing amelanotic mouse melanoma was used in conjunction with adoptively transferred OT-1+ cytotoxic T-cells expressing GFP to image antigen-specific live T-cell behavior within the tumor microenvironment. Dynamic image analysis of T-cell motility showed distinct CD8+ T-cell migration patterns and morpho-dynamics within different tumor compartments in response to ICIs: this approach was used to cluster T-cell behavior into four groups based on velocity and meandering index. The results showed that most T-cells within the tumor periphery demonstrated Lévy-like trajectories, consistent with tumor cell searching strategies. T-cells adjacent to tumor cells had reduced velocity and appeared to probe the local environment, consistent with cell-cell interactions. An increased number of T-cells were detected following treatment, traveling at lower mean velocities than controls, and demonstrating reduced displacement consistent with target engagement. Histogram-based analysis of immunofluorescent images from harvested tumors showed that in the ICI-treated mice there was a higher density of CD31+ vessels compared to untreated controls and a greater infiltration of T-cells towards the tumor core, consistent with increased cellular trafficking post-treatment

HUBUNGAN INSOMNIA TERHADAP KONSENTRASI BELAJAR MAHASISWA FAKULTAS KEDOKTERAN DI KUPANG

Seseorang yang mengalami insomnia akan mengantuk pada siang hari, sehingga dapat menurunkan konsentrasi dan akan mengganggu aktivitas. Konsentrasi diperlukan agar dapat berpikir dan bertindak untuk memiliki perhatian terhadap objek yang dipelajari dengan mengesampingkan hal-hal lain yang tidak ada hubungannya dengan pembelajaran. Tujuan penelitian ini untuk mengetahui hubungan insomnia terhadap konsentrasi belajar mahasiswa Fakultas Kedokteran di Kupang. Metode yang digunakan dalam penelitian ini jenis penelitian kuntitatif. Desain penelitian ini adalah observasional analitik dengan pendekatan cross sectional. Teknik pengambilan sampel dalam penelitian ini adalah proportional stratified random sampling dengan jumlah sampel sebanyak 76 orang. Pengumpulan data menggunakan kuesioner KSPBJ-IRS untuk mengukur insomnia dan kuesioner Konsentrasi Belajar untuk mengukur kemampuan konsentrasi belajar. Analisis data yang digunakan adalah uji spearman rank. Hasil analisis bivariat pada 76 orang mahasiswa menunjukkan adanya hubungan signifikan (p=0,000) antara insomnia dengan konsentrasi belajar. Kesimpulan penelitian ini  terdapat hubungan antara insomnia dengan konsentrasi belajar pada mahasiswa Fakultas Kedokteran di Kupang

Inducible localized delivery of an anti-PD-1 scFv enhances anti-tumor activity of ROR1 CAR-T cells in TNBC

BACKGROUND: Chimeric antigen receptor (CAR)-T cells can induce powerful immune responses in patients with hematological malignancies but have had limited success against solid tumors. This is in part due to the immunosuppressive tumor microenvironment (TME) which limits the activity of tumor-infiltrating lymphocytes (TILs) including CAR-T cells. We have developed a next-generation armored CAR (F i-CAR) targeting receptor tyrosine kinase-like orphan receptor 1 (ROR1), which is expressed at high levels in a range of aggressive tumors including poorly prognostic triple-negative breast cancer (TNBC). The F i-CAR-T is designed to release an anti-PD-1 checkpoint inhibitor upon CAR-T cell activation within the TME, facilitating activation of CAR-T cells and TILs while limiting toxicity. METHODS: To bolster potency, we developed a F i-CAR construct capable of IL-2-mediated, NFAT-induced secretion of anti-PD-1 single-chain variable fragments (scFv) within the tumor microenvironment, following ROR1-mediated activation. Cytotoxic responses against TNBC cell lines as well as levels and binding functionality of released payload were analyzed in vitro by ELISA and flow cytometry. In vivo assessment of potency of F i-CAR-T cells was performed in a TNBC NSG mouse model. RESULTS: F i-CAR-T cells released measurable levels of anti-PD-1 payload with 5 h of binding to ROR1 on tumor and enhanced the cytotoxic effects at challenging 1:10 E:T ratios. Treatment of established PDL1 + TNBC xenograft model with F i-CAR-T cells resulted in significant abrogation in tumor growth and improved survival of mice (71 days), compared to non-armored CAR cells targeting ROR1 (F CAR-T) alone (49 days) or in combination with systemically administered anti-PD-1 antibody (57 days). Crucially, a threefold increase in tumor-infiltrating T cells was observed with F i-CAR-T cells and was associated with increased expression of genes related to cytotoxicity, migration and proliferation. CONCLUSIONS: Our next-generation of ROR1-targeting inducible armored CAR platform enables the release of an immune stimulating payload only in the presence of target tumor cells, enhancing the therapeutic activity of the CAR-T cells. This technology provided a significant survival advantage in TNBC xenograft models. This coupled with its potential safety attributes merits further clinical evaluation of this approach in TNBC patients

Danish study of Non-Invasive testing in Coronary Artery Disease (Dan-NICAD):study protocol for a randomised controlled trial

BACKGROUND: Coronary computed tomography angiography (CCTA) is an established method for ruling out coronary artery disease (CAD). Most patients referred for CCTA do not have CAD and only approximately 20–30 % of patients are subsequently referred to further testing by invasive coronary angiography (ICA) or non-invasive perfusion evaluation due to suspected obstructive CAD. In cases with severe calcifications, a discrepancy between CCTA and ICA often occurs, leading to the well-described, low-diagnostic specificity of CCTA. As ICA is cost consuming and involves a risk of complications, an optimized algorithm would be valuable and could decrease the number of ICAs that do not lead to revascularization. The primary objective of the Dan-NICAD study is to determine the diagnostic accuracy of cardiac magnetic resonance imaging (CMRI) and myocardial perfusion scintigraphy (MPS) as secondary tests after a primary CCTA where CAD could not be ruled out. The secondary objective includes an evaluation of the diagnostic precision of an acoustic technology that analyses the sound of coronary blood flow. It may potentially provide better stratification prior to CCTA than clinical risk stratification scores alone. METHODS/DESIGN: Dan-NICAD is a multi-centre, randomised, cross-sectional trial, which will include approximately 2,000 patients without known CAD, who were referred to CCTA due to a history of symptoms suggestive of CAD and a low-risk to intermediate-risk profile, as evaluated by a cardiologist. Patient interview, sound recordings, and blood samples are obtained in connection with the CCTA. All patients with suspected obstructive CAD by CCTA are randomised to either stress CMRI or stress MPS, followed by ICA with fractional flow reserve (FFR) measurements. Obstructive CAD is defined as an FFR below 0.80 or as high-grade stenosis (>90 % diameter stenosis) by visual assessment. Diagnostic performance is evaluated as sensitivity, specificity, predictive values, likelihood ratios, and C statistics. Enrolment commenced in September 2014 and is expected to be complete in May 2016. DISCUSSION: Dan-NICAD is designed to assess whether a secondary perfusion examination after CCTA could safely reduce the number of ICAs where revascularization is not required. The results are expected to add knowledge about the optimal algorithm for diagnosing CAD. TRIAL REGISTRATION: Clinicaltrials.gov identifier, NCT02264717. Registered on 26 September 2014. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13063-016-1388-z) contains supplementary material, which is available to authorized users

Ross River virus antibody prevalence in the Fiji Islands, 2013-2015

A unique outbreak of Ross River virus (RRV) infection was reported in Fiji in 1979. In 2013, 29 RRV seroprevalence among residents was 46.5%. Of those born after 1982, 37.4% had anti-RRV 30 antibodies. Between 2013-2015, 10.9% of residents had seroconverted to RRV suggesting 31 ongoing endemic circulation of RRV in Fiji