Intra-tumoral heterogeneity of tumour potential doubling times (Tpot) in colorectal cancer.

Intra-tumoural heterogeneity of proliferation has been assessed by taking multiple biopsies from 30 colorectal cancers. Following in vivo IUDR labelling, dual parameter flow cytometry was used to measure tumour DNA index (DI) and labelling index (LI) and to derive DNA synthesis time (Ts) and potential doubling time (Tpot). Heterogeneity was seen for all parameters under investigation. Overall coefficients of variation (CV) and logarithmic transformation of Ts and Tpot (due to their non-gaussian distributions) indicate that LI (CV 25%) was the most variable parameter. Intra-tumoral heterogeneity in Tpot (lnTpot CV = 22%) was less than inter-individual variation (CV = 63%), suggesting that this variation should not be a limitation to the possible usefulness of this technique as an independent prognostic indicator. Correlations of Tpot values were examined between the shortest, the median and the value for a pooled homogenate sample from a single tumour. Using an homogenate, it was possible to accurately predict classification of tumour Tpot values as being below the median ('fast tumours') in 15 of 19 cases (79%). The data suggest that assaying an homogenate may allow a more rapid analysis of a multiply sampled tumour

Macrophage activation and human immunodeficiency virus infection: HIV replication directs macrophages towards a pro-inflammatory phenotype while previous activation modulates macrophage susceptibility to infection and viral production

AbstractMacrophages are pivotal for the regulation of immune and inflammatory responses, but whether their role in HIV infection is protective or deleterious remains unclear. In this study, we investigated the effect of pro- and anti-inflammatory stimuli on macrophage sensitivity to two different aspects of HIV infection: their susceptibility to infection stricto sensu, which we measured by endpoint titration method, and their ability to support virus spread, which we measured by using an RT activity assay in infection kinetics. We show a partially protective role for pro-inflammatory agents as well as for IL-4. We also illustrate that various different stimuli display differential effects on macrophage susceptibility to HIV and on virus replication that occurs thereafter. On the other hand, HIV replication strongly repressed CD206 and CD163 expression, thus clearly orientating macrophages towards a pro-inflammatory phenotype, but independently of TNF. Taken together, our results emphasize that HIV infection of macrophages sets up inflammation at the cell level but through unexpected mechanisms. This may limit target susceptibility and participate in virus clearance but may also result in tissue damage

Potential worldwide impacts of sea level rise on coastal-lowland anurans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Amphibians are the most severely threatened terrestrial vertebrates and we are witnessing a global decline phenomenon, which is even suggested to be of the same level as the historical mass extinctions. Albeit the myriad of causative stressors identified in the last decades, future sea level rise (SLR) and its impact on coastal terrestrial fauna remains essentially unreported. Even if there is no consensus on the magnitude of the future SLR, several studies suggest that it is likely to be greater than previously reported by the Fourth Assessment Report (AR4) of the Intergovernmental Panel on Climate Change (IPCC). Therefore, it is reasonable to expect severe impacts on the coastal terrestrial fauna at a worldwide scale. Here, we assembled a worldwide data set of coastal-lowland anuran species in an attempt to quantify the potential habitat loss caused by flooding according to different SLR scenarios. We also assessed potential habitat suitability under climate change (CC) in order to evaluate its expected effects on species' climatic niches, by building species distribution models for three future scenarios (A2a, A1b and B2a). Our results revealed that SLR has the potential to produce negative impacts on similar to 86% of the selected coastal-lowland species in different magnitudes, whereas CC is expected to produce a greater impact on the same taxa. Thus, species predicted to persist under the new climatic conditions may be exposed to effects associated with SLR. Breaking our results down to biogeographic realms, we found that Australasia harboured most amphibian species suffering the dual impacts of SLR and CC. Based on our results, we advocate for the inclusion of potential future impacts of SLR in conservation action plans, anticipating and preventing biodiversity loss.Amphibians are the most severely threatened terrestrial vertebrates and we are witnessing a global decline phenomenon, which is even suggested to be of the same level as the historical mass extinctions. Albeit the myriad of causative stressors identified12191101CNQP - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)161812/2011-23855/2013- 92011/51694-7405285/2013-2302589/2013-9We thank Rafael Loyola, Jean Vitule, Faraham Ahmadzadeh, Adriele Oliveira, and Guilherme Becker for their suggestions and contributions. ISO is grateful to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq: 161812/2011-2) and Coordenaçã

An immunohistochemical assessment of cellular proliferation markers in head and neck squamous cell cancers.

Prognostic information is essential for the evaluation, judgement and optimal treatment of patients with squamous cell cancers (SCCs) of the upper aerodigestive tract. Using immunohistochemical and flow cytometric techniques, we have studied the significance of cellular expression of the Ki-67 antigen, epidermal growth factor receptor (EGFR), the transferrin receptor (TFR) and DNA ploidy status in a prospective analysis of patients with SCCs of the head and neck region. All 42 fresh tumour samples (five well differentiated; 28 moderately differentiated; nine poorly differentiated) expressed both EGFR and TFR to varying degrees. Receptor expression was most marked on the peripheral invading margin of cancer cell islands although staining was also demonstrated in a random fashion within cellular islands and consistently along the basal cell layer of overlying stratified squamous epithelium. The percentage of cancer cells that reacted with the Ki-67 monoclonal antibody was assessed as low (less than 10%) in 15 samples (35.8%), intermediate (10-30%) in 19 samples (45.2%) and high (greater than 30%) in eight samples (19.0%). Eleven of 15 samples (73%) with a low percentage reactivity were DNA diploid, whereas seven of eight samples (87.5%) with a high percentage reactivity were DNA aneuploid. Poorly differentiated SCCs were significantly more often aneuploid than were either moderately or well differentiated tumours. Our results suggest that EGFR and TFR are widely distributed on SCCs, especially on proliferating cells at the invading tumour margin. In addition, there is a close spatial correlation between cells expressing EGFR, TFR and those expressing the Ki-67 antigen. Tumours in which the staining intensity for both EGFR and TFR was intense invariably expressed the Ki-67 antigen in a high proportion of cells. Further patient follow-up will be important in determining whether intense EGFR and TFR staining, combined with a high percentage reactivity with Ki-67 antibody and DNA aneuploidy, will ultimately define a subset of head and neck cancer patients with a poor clinical outcome

A Plant-Made Vaccine Candidate To Protect Ruminants Against Shiga Toxin-Producing Escherichia Coli

Shiga toxin-producing Escherichia coli (STEC) are enteropathogens colonizing the digestive tracts of humans and animals worldwide. STEC are shed in the manure of cattle and frequently contaminate beef products, vegetables, and drinking water. Immunizing cattle herds against STEC is a promising strategy to reduce STEC colonization in cattle and therefore decrease contamination in the food supply. The goal of this project is to produce a plant-made vaccine to protect ruminants against STEC. Several recombinant proteins were designed as vaccine candidates and expressed transiently in Nicotiana benthamiana and transplastomically Nicotiana tabacum. One vaccine candidate, an E. coli secreted protein A (EspA) fusion, accumulated in transplastomic lines up to 220.7 mg/kg of fresh leaf weight. Leaf tissue containing the antigen was lyophilized and will be orally administered to a small ruminant model to test vaccine efficacy

Altimetry for the future: Building on 25 years of progress

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Altimetry for the future: Building on 25 years of progress

In 2018 we celebrated 25 years of development of radar altimetry, and the progress achieved by this methodology in the fields of global and coastal oceanography, hydrology, geodesy and cryospheric sciences. Many symbolic major events have celebrated these developments, e.g., in Venice, Italy, the 15th (2006) and 20th (2012) years of progress and more recently, in 2018, in Ponta Delgada, Portugal, 25 Years of Progress in Radar Altimetry. On this latter occasion it was decided to collect contributions of scientists, engineers and managers involved in the worldwide altimetry community to depict the state of altimetry and propose recommendations for the altimetry of the future. This paper summarizes contributions and recommendations that were collected and provides guidance for future mission design, research activities, and sustainable operational radar altimetry data exploitation. Recommendations provided are fundamental for optimizing further scientific and operational advances of oceanographic observations by altimetry, including requirements for spatial and temporal resolution of altimetric measurements, their accuracy and continuity. There are also new challenges and new openings mentioned in the paper that are particularly crucial for observations at higher latitudes, for coastal oceanography, for cryospheric studies and for hydrology. The paper starts with a general introduction followed by a section on Earth System Science including Ocean Dynamics, Sea Level, the Coastal Ocean, Hydrology, the Cryosphere and Polar Oceans and the “Green” Ocean, extending the frontier from biogeochemistry to marine ecology. Applications are described in a subsequent section, which covers Operational Oceanography, Weather, Hurricane Wave and Wind Forecasting, Climate projection. Instruments’ development and satellite missions’ evolutions are described in a fourth section. A fifth section covers the key observations that altimeters provide and their potential complements, from other Earth observation measurements to in situ data. Section 6 identifies the data and methods and provides some accuracy and resolution requirements for the wet tropospheric correction, the orbit and other geodetic requirements, the Mean Sea Surface, Geoid and Mean Dynamic Topography, Calibration and Validation, data accuracy, data access and handling (including the DUACS system). Section 7 brings a transversal view on scales, integration, artificial intelligence, and capacity building (education and training). Section 8 reviews the programmatic issues followed by a conclusion