Dynamics of Adrenal Steroids Are Related to Variations in Th1 and Treg Populations during Mycobacterium tuberculosis Infection in HIV Positive Persons

Tuberculosis (TB) remains the most frequent cause of illness and death from an infectious agent, and its interaction with HIV has devastating effects. We determined plasma levels of dehydroepiandrosterone (DHEA), its circulating form DHEA-suphate (DHEA-s) and cortisol in different stages of M. tuberculosis infection, and explored their role on the Th1 and Treg populations during different scenarios of HIV-TB coinfection, including the immune reconstitution inflammatory syndrome (IRIS), a condition related to antiretroviral treatment. DHEA levels were diminished in HIV-TB and HIV-TB IRIS patients compared to healthy donors (HD), HIV+ individuals and HIV+ individuals with latent TB (HIV-LTB), whereas dehydroepiandrosterone sulfate (DHEA-s) levels were markedly diminished in HIV-TB IRIS individuals. HIV-TB and IRIS patients presented a cortisol/DHEA ratio significantly higher than HIV+, HIV-LTB and HD individuals. A positive correlation was observed between DHEA-s and CD4 count among HIV-TB individuals. Conversely, cortisol plasma level inversely correlated with CD4 count within HIV-TB individuals. M. tuberculosis-specific Th1 lymphocyte count was increased after culturing PBMC from HIV-TB individuals in presence of DHEA. We observed an inverse correlation between DHEA-s plasma level and Treg frequency in co-infected individuals, and CD4+FoxP3+ Treg frequency was increased in HIV-TB and IRIS patients compared to other groups. Strikingly, we observed a prominent CD4+CD25-FoxP3+ population across HIV-TB and HIV-TB IRIS patients, which frequency correlated with DHEA plasma level. Finally, DHEA treatment negatively regulated FoxP3 expression without altering Treg frequency in co-infected patients. These data suggest an enhancing role for DHEA in the immune response against M. tuberculosis during HIV-TB coinfection and IRIS

Joint Tumor Bud–MMP/TIMP Count at the Invasive Front Improves the Prognostic Evaluation of Invasive Breast Carcinoma

Background: Tumor budding is a histological phenomenon consisting of the formation of small clusters of one to five undifferentiated malignant cells detached from the main tumor mass which are observed in the tumor stroma. In the present study, we investigated the prognostic significance of tumor budding in breast cancer and its relationship with the expressions of matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs). Methods: The number of buds was counted in whole-tissue sections from 153 patients with invasive ductal carcinomas who underwent a long follow-up period. In addition, an immunohistochemical study of MMP-9, -11, and -14 TIMP-1 and -2 expression by cell types at the invasive tumor front was carried out. Results: There was a wide variability in the number of buds among tumors, ranging from 0 to 28 (median = 5). Tumor budding count ≥ 4 was the optimal cut-off to predict both relapse-free and overall survival. High-grade tumor budding was associated with MMP/TIMP expression by cancer-associated fibroblasts. In addition, we found that the combination of tumor budding grade with MMP/TIMP expression by stromal cells, and especially with MMP-11 expression by mononuclear inflammatory cells, significantly improved the prognostic evaluation. Conclusion: High-grade tumor budding is associated with a more aggressive tumor phenotype, which, combined with MMP/TIMP expression by stromal cells at the invasive front of the tumor, identifies patients with poor prognosis

Epidemiological characteristics of the subjects enrolled.

<p>BCG, Bacillus Calmette-Guerin. IQR, Interquartile range.</p

DHEA modulates the expression of the FoxP3 transcription factor in coinfected individuals. A and B.

<p>Percentages of Treg cells (defined as CD4+FoxP3+CD25+) in PBMC from <b>A.</b> HIV-TB and <b>B.</b> HD individuals stimulated with <i>M. tuberculosis</i> antigen in the presence or absence of DHEA at the indicated concentrations for 3 days. Bars indicate the mean ± SEM for each experimental condition. <b>C and D.</b> Relative FoxP3 Median florescence intensity (MFI) in Treg lymphocytes from <b>C.</b> HIV-TB patients and <b>D.</b> HD volunteers after culturing PBMC as detailed in A. Bars indicate the mean ± SEM for each treatment. Data are representative of four different experiments. *: <i>p</i> < 0.05; **: <i>p</i> < 0.01. <b>E.</b> Representative flow cytometry graphs depicting the results obtained from culturing PBMC from HIV-TB and HD individuals as indicated above.</p

Modulation of <i>M. tuberculosis</i>-induced IFN-γ production by adrenal hormones.

<p><b>A.</b> Absolute numbers of IFN-γ producer cells from PBMC of HIV+, HIV-TB, IRIS, HIV-LTB and HD patients, which have been stimulated with <i>M. tuberculosis</i> antigen for 16 hours. Horizontal lines indicate the mean and comparisons between groups and statistically significant differences are shown. SFU, Spots forming units. <b>B.</b> Percentage of IFN-γ producer cells relative to <i>M. tuberculosis</i> in PBMC of HIV-TB patients (n = 12). PBMC (10⁵ cells/well) were stimulated in the presence of <i>M. tuberculosis</i> with or without addition of DHEA and/or cortisol at the indicated concentrations. Each bar illustrates the mean ± SEM of the percentage for IFN-γ producer cells relative to <i>M. tuberculosis</i> for the each group, calculated as follows: % of IFN-γ relative to <i>Mtb</i> = ([(<i>Mtb</i> hormone-Media)-(<i>Mtb</i>-Media)]/(<i>Mtb</i>-Media))×100. Asterisks indicate comparisons between each condition against <i>Mtb</i> specific response. *: <i>p</i> < 0.05; **: <i>p</i> < 0.01.</p

Expanded “non-conventional” Treg in HIV-TB and IRIS patients.

<p>Analysis of CD4+FoxP3+CD25+ and CD4+FoxP3+CD25- populations within Treg in <b>A;</b> HIV-TB (n = 11), <b>B;</b> IRIS (n = 5), <b>C;</b> HIV-LTB (n = 4), <b>D;</b> HD (n = 10) and <b>E;</b> HIV (n = 7) individuals. Horizontal lines inside the boxes indicate the means ± SEM of each group. Asterisks denote comparisons between each sub-population. *: p < 0.05; **: p < 0.01; ***: p< 0.001. <b>F.</b> Spearman correlation analysis between plasma DHEA and the percentage of CD4+FoxP3+CD25- lymphocytes from all grouped patients (HIV+, HIV-TB, IRIS, HIV-LTB and HD). Results of statistical analysis are shown in the graphic.</p

Cortisol, dehidroepindrosterone-sulfate (DHEA-s), DHEA levels and Cortisol/DHEA ratio in HIV, HIV-TB, IRIS, HIV-LTB and HD.

<p><b>A.</b> DHEA, <b>B.</b> DHEA-s, <b>C.</b> Cortisol and <b>D.</b> Cortisol/DHEA (ratio) levels in plasma of HIV+, HIV-TB, IRIS, HIV-LTB, and HD individuals. Bars indicate the mean ± SEM for each group. Horizontal lines indicate comparisons between groups and statistically significant differences. DHEA was measured by radioimmunoassay, DHEA-s by immunochemoluminiscence tests and Cortisol by electrochemiluminescence. HIV+ individuals n = 10, HIV-TB n = 21, IRIS n = 6, HIV-LTB n = 5, and HD n = 16. *: <i>p</i> < 0.05; **: <i>p</i> < 0.01; ***: <i>p</i> < 0.001.</p

Assessment of Mediterranean Aquaculture Sustainability

Aquaculture is a source of food, critical and essential to feed humanity and to ensure the world’s food security, and, also is a business that generates economic interest. This is clearly specified in the analysis document of the State of Fisheries and Aquaculture FAO 2016, which exposes that “aquaculture will become the main driver of change in the fisheries and aquaculture sector“. Aquaculture is the productive industrial activity that will play a crucial role in providing solutions to the millennium challenges. Globally this is the main idea that exists under the MedAID EU Horizon 2020 project. Production and productivity of Mediterranean marine fish aquaculture, mainly seabass and seabream, are stagnating or growing slowly as a result of multiple and interrelated causes. To accomplish the objective of improving its competitiveness and sustainability, MedAID is structured in a first interdisciplinary Work-packages (WPs) to assess technical, environmental, market, socioeconomic and governance weaknesses. WP1, “Holistic sustainability assessment of Mediterranean marine fish farming sector”, aims to carry out this mentioned assessment with a particular focus on the farm’s zootechnical performance, environmental and social sustainability, health, welfare and diseases, economic aspects and governance. The objective is to provide an overview of all the components of the value chain and assist the other WPs in their execution. To succeed in this challenge we started from the knowledge that we get from surveys addressed to the sector. The aim of the present report “Deliverable 1.2. Assessment of Mediterranean Aquaculture Sustainability” is to present the results of such assessment throughout the compilation of different analysis (Zotechnical performance, Environmental and social sustainability, Prevalence of diseases, Economic performance of the industry, and Governance and social acceptability), which has been mainly done based on a survey addressed to Mediterranean aquaculture farms from the whole region. The replies to surveys were compiled in a the Database of Mediterranean aquaculture farms, which have been used to analyse the information from a wider perspective, for both technical and economic indicators considered in the different thematic analysis here presented. MedAID has been able to obtain information from 27 partners and collaborating companies (50 production units) from 10 countries. For confidentially reasons their names are no mentioned in this report. Our most sincere gratitude is due to them, as without their collaboration, this study would had not been possible. Although not all companies replied to all questions, and for some parts of the analysis the data were small, this study was able to identify significant associations between key Performance Indicators (KPIs) and predictor variables in the units that took part in the survey. The survey data analysis shows that farming practices differ among the farms, allowing for improvement of management and, also, there is not a common production method for the Mediterranean seabass and seabream production. Moreover, it should be considered both seabass and seabream routinely surveyed and analysed separately, and the possibility of analysing separately the farms according production systems. Companies need to improve their productive efficiency through technical, operational and management innovations with a real transfer to the productive activity. The reduction of the average cost of production is a key aspect for the sustainability of the activity. While diversification is more efficient as the size of the company increases. Finally, there is a broad consensus about the necessity of improving governance, reducing the complexity of the administration and the regulation of the sector. It is also argued the need to reduce the time required to obtain a license