Variants in the inflammatory IL6 and MPO genes modulate stroke susceptibility through main effects and gene-gene interactions

A complex interplay between genetic background, clinical and life-style factors and the environment is expected to ultimately regulate the onset, acute phase and outcome of stroke. There is substantial evidence that inflammation within the Central Nervous System contributes to stroke risk, and known clinical risk factors for stroke, like atherosclerosis, diabetes, obesity, hypertension, and peripheral infection, are associated with an elevated systemic inflammatory profile. The inflammatory response is equally of major importance in recovery and healing processes after stroke. In this study we tested the genetic association of major inflammatory players IL1B (2q14), IL6 (7p21), TNF (6p21.3) and MPO (17q23.1) with stroke susceptibility and with stroke outcome at three months, in a population sample of 672 patients and 530 controls, adjusting for demographic, clinical and life-style risk factors and/or stroke severity parameters. The apparent complexity of the inflammatory mechanisms in stroke, and the multiplicity of players involved suggest a concerted process, in which implicated molecules interact to tightly regulate each other. We therefore examined both independent gene effects and the occurrence of gene-gene interactions among the tested inflammatory genes in stroke risk and stroke recovery. Two IL6 and one MPO SNP were significantly associated with stroke risk after multiple testing correction (0.022 correctedP 0.042), highlighting gene variants of low to moderate effect in stroke risk. An epistatic interaction between the IL6 and MPO genes was also identified in association with stroke susceptibility (P=0.031 after 1000 permutations). In the subset of 546 patients assessed for stroke outcome at three months using the modified Rankin Scale (mRS), we found one IL6 haplotype associated with stroke outcome (correctedP=0.024). In the present study we present supporting evidence for a role of the IL6 and MPO inflammatory genes in stroke susceptibility, and show that stroke risk is modulated by main gene effects together with clinical and life-style factors as well as by gene-gene interactions. Our findings are compatible and strengthen previous genetic and biological observations, highlighting the need of further functional studies, particularly in view of the possible utility of IL-6 as a diagnostic and/or prognostic biomarker for stroke

Patterns of colonization and species distribution for Azorean arthropods: evolution, diversity, rarity and extinction

Proceedings of the Symposium "Darwin's Mistake and what we are doing to correct it". Ponta Delgada, 19-22 September, 2009.Com base em estudos ecológicos e biogeográficos realizados nos Açores (um arquipélago remoto composto por nove ilhas vulcânicas) durante muitos anos de uma forma continuada, apresentamos um conjunto de questões. O grupo alvo são os artrópodes e o principal habitat é a Laurissilva, a floresta nativa dos Açores. A diversificação das espécies de artrópodes dos Açores é afectada pela idade das ilhas, área das ilhas e seu isolamento. No entanto, os estudos que decorreram durante os últimos dez anos mostram que os vários tipos de grupos taxonómicos e ecológicos são afectados de forma diferente por estes factores, tendo como consequência padrões de distribuição espacial únicos. A distribuição espacial dos artrópodes em cada ilha é causada for "efeitos de massa", muitas espécies possuindo dinâmicas "fonte-sumidouro". As espécies verdadeiramente raras à escala regional são aquelas que são especialistas de um particular habitat, muitas delas sendo espécies endémicas ameaçadas. Embora várias espécies endémicas persistam com populações sumidouro em habitats criados pelo Homem (e.g. florestas exóticas), mais de metade das espécies especialistas da floresta nativa já estão extintas ou poderão extinguir-se no futuro. De facto, aquelas espécies que evoluíram e apenas são encontradas nas florestas nativas foram afectadas de forma dramática como resultado da destruição alargada das florestas nativas dos Açores. Defendemos que a única forma de evitar a extinção de uma fauna única das florestas nativas dos Açores será através de medidas de restauro desta floresta.ABSTRACT: Here we address a list of questions based on long-term ecological and biogeographical studies performed in the Azores, a remote volcanic oceanic archipelago composed by nine islands. The target group are the arthropods, and the main habitat the Laurisilva, the Azorean native forest. Diversification of Azorean arthropod species is affected by island age, area and isolation. However, results obtained for over a decade show that distinct groups are differently affected by these factors, which has lead to the extreme diverse distribution patterns currently observed. Spatial distribution of arthropods in each island may be interpreted as caused by a typical "mass effect", with many species following a "source-sink" dynamics. Truly regionally rare species are those that are habitat specialists, many of them being threatened endemic species. Although various endemics persist as sink populations in human-made habitats (e.g., exotic forests), more than half of the original endemic forest arthropods may already have vanished or may eventually be driven to extinction in the future. Those species which have evolved in and are mainly found in native forests, have been dramatically affected by hitherto unrecognized levels of extinction debt, as a result of extensive destruction of native forest. We argue that immediate action to restore and expand native forest habitat is required to avoid a future of disastrous extinctions of a biologically unique fauna with an unique evolutionary history

High resolution melting : improvements in the genetic diagnosis of hypertrophic cardiomyopathy in a Portuguese cohort

© 2012 Susana et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background: Hypertrophic Cardiomyopathy (HCM) is a complex myocardial disorder with a recognized genetic heterogeneity. The elevated number of genes and mutations involved in HCM limits a gene-based diagnosis that should be considered of most importance for basic research and clinical medicine. Methodology: In this report, we evaluated High Resolution Melting (HRM) robustness, regarding HCM genetic testing, by means of analyzing 28 HCM-associated genes, including the most frequent 4 HCM-associated sarcomere genes, as well as 24 genes with lower reported HCM-phenotype association. We analyzed 80 Portuguese individuals with clinical phenotype of HCM allowing simultaneously a better characterization of this disease in the Portuguese population. Results: HRM technology allowed us to identify 60 mutated alleles in 72 HCM patients: 49 missense mutations, 3 nonsense mutations, one 1-bp deletion, one 5-bp deletion, one in frame 3-bp deletion, one insertion/deletion, 3 splice mutations, one 5’UTR mutation in MYH7, MYBPC3, TNNT2, TNNI3, CSRP3, MYH6 and MYL2 genes. Significantly 22 are novel gene mutations. Conclusions: HRM was proven to be a technique with high sensitivity and a low false positive ratio allowing a rapid, innovative and low cost genotyping of HCM. In a short return, HRM as a gene scanning technique could be a cost-effective gene-based diagnosis for an accurate HCM genetic diagnosis and hopefully providing new insights into genotype/phenotype correlations.info:eu-repo/semantics/publishedVersio

How can biomolecules improve mucoadhesion of oral insulin? A comprehensive insight using ex-vivo, in silico, and in vivo models

Funding: Supported in part by UID/DTP/04138/2019 from FCT, Portugal and DREAMS (ULHT). SEM analysis was funded by FCT/MCTES for the financial support to CESAM (UIDP/50017/2020+UIDB/50017/2020), through national funds.Currently, insulin can only be administered through the subcutaneous route. Due to the flaws associated with this route, it is of interest to orally deliver this drug. However, insulin delivered orally has several barriers to overcome as it is degraded by the stomach’s low pH, enzymatic content, and poor absorption in the gastrointestinal tract. Polymers with marine source like chitosan are commonly used in nanotechnology and drug delivery due to their biocompatibility and special features. This work focuses on the preparation and characterization of mucoadhesive insulin-loaded polymeric nanoparticles. Results showed a suitable mean size for oral administration (<600 nm by dynamic laser scattering), spherical shape, encapsulation efficiency (59.8%), and high recovery yield (80.6%). Circular dichroism spectroscopy demonstrated that protein retained its secondary structure after encapsulation. Moreover, the mucoadhesive potential of the nanoparticles was assessed in silico and the results, corroborated with ex-vivo experiments, showed that using chitosan strongly increases mucoadhesion. Besides, in vitro and in vivo safety assessment of the final formulation were performed, showing no toxicity. Lastly, the insulin-loaded nanoparticles were effective in reducing diabetic rats’ glycemia. Overall, the coating of insulin-loaded nanoparticles with chitosan represents a potentially safe and promising approach to protect insulin and enhance peroral delivery.publishersversionpublishe

A Step Forward in Breast Cancer Research: From a Natural-Like Experimental Model to a Preliminary Photothermal Approach

Supplementary Materials - The following are available online at https://www.mdpi.com/1422-0067/21/24/9681/s1, Figure S1: GNPs’ size distribution by intensity (%) obtained by DLS.Breast cancer is one of the most frequently diagnosed malignancies and common causes of cancer death in women. Recent studies suggest that environmental exposures to certain chemicals, such as 7,12-Dimethylbenzanthracene (DMBA), a chemical present in tobacco, may increase the risk of developing breast cancer later in life. The first-line treatments for breast cancer (surgery, chemotherapy or a combination of both) are generally invasive and frequently associated with severe side effects and high comorbidity. Consequently, novel approaches are strongly required to find more natural-like experimental models that better reflect the tumors’ etiology, physiopathology and response to treatments, as well as to find more targeted, efficient and minimally invasive treatments. This study proposes the development and an in deep biological characterization of an experimental model using DMBA-tumor-induction in Sprague-Dawley female rats. Moreover, a photothermal therapy approach using a near-infrared laser coupled with gold nanoparticles was preliminarily assessed. The gold nanoparticles were functionalized with Epidermal Growth Factor, and their physicochemical properties and in vitro effects were characterized. DMBA proved to be a very good and selective inductor of breast cancer, with 100% incidence and inducing an average of 4.7 tumors per animal. Epigenetic analysis showed that tumors classified with worst prognosis were hypomethylated. The tumor-induced rats were then subjected to a preliminary treatment using functionalized gold nanoparticles and its activation by laser (650–900 nm). The treatment outcomes presented very promising alterations in terms of tumor histology, confirming the presence of necrosis in most of the cases. Although this study revealed encouraging results as a breast cancer therapy, it is important to define tumor eligibility and specific efficiency criteria to further assess its application in breast cancer treatment on other species.The APC was funded by Faculty of Pharmacy, University of Coimbra and Coimbra Chemistry Centre, Department of Chemistry, University of Coimbra. This work was also supported by Fundação para a Ciência e Tecnologia (FCT), Portugal under the project’s references UIDB/00645/2020 and UID/DTP/04138/2019. TFG was supported by FCT, Portugal under the reference SFRH/BD/147306/2019. Thanks to FCT/MCTES for the financial support to CESAM (UIDP/50017/2020+UIDB/50017/2020), through national funds.info:eu-repo/semantics/publishedVersio

Customized tracheal design using 3D printing of a polymer hydrogel: influence of UV laser cross-linking on mechanical properties

H2020-ICT-2014-1,TransFlexTeg645241 ERC-CoG-2014, CapTherPV, 647596 Pest-UID/FIS/00068/2013The use of 3D printing of hydrogels as a cell support in bio-printing of cartilage, organs and tissue has attracted much research interest. For cartilage applications, hydrogels as soft materials must show some degree of rigidity, which can be achieved by photo- or chemical polymerization. In this work, we combined chemical and UV laser polymeric cross-linkage to control the mechanical properties of 3D printed hydrogel blends. Since there are few studies on UV laser cross-linking combined with 3D printing of hydrogels, the work here reported offered many challenges.publishe

Effects of recent minimum temperature and water deficit increases on Pinus pinaster radial growth and wood density in southern Portugal

Western Iberia has recently shown increasing frequency of drought conditions coupled with heatwave events, leading to exacerbated limiting climatic conditions for plant growth. It is not clear to what extent wood growth and density of agroforestry species have suffered from such changes or recent extreme climate events. To address this question, tree-ring width and density chronologies were built for a Pinus pinaster stand in southern Portugal and correlated with climate variables, including the minimum, mean and maximum temperatures and the number of cold days. Monthly and maximum daily precipitations were also analyzed as well as dry spells. The drought effect was assessed using the standardized precipitation-evapotranspiration (SPEI) multi-scalar drought index, between 1 to 24-months. The climate-growth/density relationships were evaluated for the period 1958-2011. We show that both wood radial growth and density highly benefit from the strong decay of cold days and the increase of minimum temperature. Yet the benefits are hindered by long-term water deficit, which results in different levels of impact on wood radial growth and density. Despite of the intensification of long-term water deficit, tree-ring width appears to benefit from the minimum temperature increase, whereas the effects of long-term droughts significantly prevail on tree-ring density. Our results further highlight the dependency of the species on deep water sources after the juvenile stage. The impact of climate changes on longterm droughts and their repercussion on the shallow groundwater table and P. pinaster’s vulnerability are also discussed. This work provides relevant information for forest management in the semi-arid area of the Alentejo region of Portugal. It should ease the elaboration of mitigation strategies to assure P. pinaster’s production capacity and quality in response to more arid conditions in the near future in the regioninfo:eu-repo/semantics/publishedVersio

Metal Coordination and Biological Screening of a Schiff Base Derived from 8-Hydroxyquinoline and Benzothiazole

Designing new metallodrugs for anticancer therapy is a driving force in the scientific community. Aiming to contribute to this field, we hereby report the development of a Schiff base (H2L) derived from the condensation of 2-carbaldehyde-8-hydroxyquinoline with 2-hydrazinobenzothiazole and its complexation with transition metal ions. All compounds were characterised by analytical and spectroscopic techniques, which disclosed their structure: [Cu(HL)Cl], [Cu(HL)2], [Ni(HL)(acetate)], [Ni(HL)2], [Ru(HL)Cl(DMSO)], [VO(HL)2] and [Fe(HL)2Cl(H2O)]. Different binding modes were proposed, showing the ligand’s coordination versatility. The ligand proton dissociation constants were determined, and the tested compounds showed high lipophilicity and light sensitivity. The stability of all complexes in aqueous media and their ability to bind to albumin were screened. Based on an antiproliferative in vitro screening, [Ni(HL)(acetate)] and [Ru(HL)Cl(DMSO)] were selected for further studies aiming to investigate their mechanisms of action and therapeutic potential towards colon cancer. The complexes displayed IC50 < 21 μM towards murine (CT-26) and human (HCT-116) colon cancer cell lines. Importantly, both complexes exhibited superior antiproliferative properties compared to the clinically approved 5-fluorouracil. [Ni(HL)(acetate)] induced cell cycle arrest in S phase in CT-26 cells. For [Ru(HL)Cl(DMSO)] this effect was observed in both colon cancer cell lines. Additionally, both compounds significantly inhibited cell migration particularly in the human colon cancer cell line, HCT-116. Overall, the therapeutic potential of both metal complexes was demonstrated

Proof-of-Concept Study of Multifunctional Hybrid Nanoparticle System Combined with NIR Laser Irradiation for the Treatment of Melanoma

Supplementary Materials - available online at https://www.mdpi.com/article/10.3390/biom11040511/s1The global impact of cancer emphasizes the importance of developing innovative, effective and minimally invasive therapies. In the context of superficial cancers, the development of a multifunctional nanoparticle-based system and its in vitro and in vivo safety and efficacy characterization are, herein, proposed as a proof-of-concept. This multifunctional system consists of gold nanoparticles coated with hyaluronic and oleic acids, and functionalized with epidermal growth factor for greater specificity towards cutaneous melanoma cells. This nanoparticle system is activated by a near-infrared laser. The characterization of this nanoparticle system included several phases, with in vitro assays being firstly performed to assess the safety of gold nanoparticles without laser irradiation. Then, hairless immunocompromised mice were selected for a xenograft model upon inoculation of A375 human melanoma cells. Treatment with near-infrared laser irradiation for five minutes combined with in situ administration of the nanoparticles showed a tumor volume reduction of approximately 80% and, in some cases, led to the formation of several necrotic foci, observed histologically. No significant skin erythema at the irradiation zone was verified, nor other harmful effects on the excised organs. In conclusion, these assays suggest that this system is safe and shows promising results for the treatment of superficial melanoma.The authors would like to thank to Fundação para a Ciência e Tecnologia (FCT) for the essential financial support under the project’s references PTDC/BBB-BMC/0611/2012, UIDB/00645/2020, UIDB/04138/2020 and UIDP/04138/2020 as well as for the PhD fellowships SFRH/BD/148044/2019 and SFRH/BD/147306/2019.info:eu-repo/semantics/publishedVersio