Design of an antifungal surface embedding liposomal Amphotericin b through a mussel adhesive-inspired coating strategy

Microbial colonisation of urinary catheters remains a serious problem for medicine as it often leads to biofilm formation and infection. Among the approaches reported to deal with this problem, surfaces functionalization to render them with antimicrobial characteristics, comprises the most promising one. Most of these strategies, however, are designed to target bacterial biofilms, while fungal biofilms are much less taken into account. In real-life settings, fungi will be inevitably found in consortium with bacteria, especially in the field of biomaterials. The development of antifungal coating strategies to be combined with antibacterial approaches will be pivotal for the fight of biomaterial-associated infections. The main goal of the present study was, therefore, to engineer an effective strategy for the immobilization of liposomal amphotericin B (LAmB) on polydimethylsiloxane (PDMS) surfaces to prevent Candida albicans colonization. Immobilization was performed using a two-step mussel-inspired coating strategy, in which PDMS are first immersed in dopamine solution. Its self-polymerization leads to the deposition of a thin adherent film, called polydopamine (pDA), which allowed the incorporation of LAmB, afterwards. Different concentrations of LAmB were screened in order to obtain a contact-killing surface with no release of LAmB. Surface characterization confirmed the polymerization of dopamine and further functionalization with LAmB yielded surfaces with less roughness and more hydrophilic features. The proposed coating strategy rendered the surfaces of PDMS with the ability to prevent the attachment of Candida albicans and kill the adherent cells, without toxicity towards mammalian cells. Overall results showed that LAmB immobilization on a surface retained its antifungal activity and reduced toxicity, holding therefore a great potential to be applied for the design of urinary catheters. Since the sessile communities commonly found associated to these devices exhibit a polymicrobial nature, the next challenge will be to co-immobilize LAmB with antibacterial agents to prevent the establishment of catheter urinary-associated infections.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The authors also acknowledge the support, through the Programa Operacional Competitividade e Internacionalizacao (COMPETE2020) and by national funds, through the Portuguese Foundation for Science and Technology (FCT), of the POLY-PrevEnTT project (PTDC/BTM-SAL/29841/2017-POCI-01-0145-FEDER-029841).info:eu-repo/semantics/publishedVersio

Modulating an antimicrobial release approach by dopamine chemistryto fight infections associated to orthopedic implants

Alongside with orthopedic implants contribution for modern healthcare improvements, theres the risk associated to their microbial colonization and biofilm formation, compromising the performance of the implant itself and representing niches for infection. This study aimed to engineer an antimicrobial release coating for stainless steel surfaces (SS) to empower them with the ability to prevent Staphylococci colonization. Surface modification was based on dopamine chemistry, which self-polymerization results in the deposition of a thin, adhered film called polydopamine (pDA). Chlorohexidine (CHX) was chosen to confer the antimicrobial features. Its immobilization was performed through a 2-step approach, including pDA formation and immersion in CHX solution, and 1-step strategy, in which dopamine and CHX were dissolved together and SS coupons were immersed in this solution. An additional layer of pDA was also performed for both strategies. SEM and AFM confirmed pDA coating by the presence of self-polymerized pDA particles without altering the roughness of SS surfaces. Immobilization of CHX using a 1-step approach yielded surfaces with a more homogenous coating than the 2-step approach. Different pDA-based strategies yielded different CHX release profiles: the amount of CHX released was higher for the 2-step approach and the addition of another pDA layer reduced the amount of CHX released. The antimicrobial performance of the modified surfaces was evaluated against S. aureus and S. epidermidis and the results showed that all the strategies caused a significant reduction (more than 3 LOG) in the number of cells adhered to the surfaces and in suspension, after 24 h. The 2-step approach was able to impart SS surfaces with antimicrobial activity even after 10 days of exposure. In conclusion, dopamine chemistry can modulate CHX release from the surfaces to obtain an antimicrobial coating strategy with great potential to fight infections associated with orthopedic implants.info:eu-repo/semantics/publishedVersio

Oriented Tailoring of Plastic Antibodies for Prostate Specific Antigen and Application of the Imprinted Material as Ionophore in Potentiometric Detection

Poster, presented at Nanobio Europe, Varese, Italy, 18 - 20 June, 2012.Prostate Specific Antigen (PSA) is the biomarker of choice for screening prostate cancer throughout the population, with PSA values above 10 ng/mL pointing out a high probability of associated cancer1. According to the most recent World Health Organization (WHO) data, prostate cancer is the commonest form of cancer in men in Europe2. Early detection of prostate cancer is thus very important and is currently made by screening PSA in men over 45 years old, combined with other alterations in serum and urine parameters. PSA is a glycoprotein with a molecular mass of approximately 32 kDa consisting of one polypeptide chain, which is produced by the secretory epithelium of human prostate. Currently, the standard methods available for PSA screening are immunoassays like Enzyme-Linked Immunoabsorbent Assay (ELISA). These methods are highly sensitive and specific for the detection of PSA, but they require expensive laboratory facilities and high qualify personal resources. Other highly sensitive and specific methods for the detection of PSA have also become available and are in its majority immunobiosensors1,3-5, relying on antibodies. Less expensive methods producing quicker responses are thus needed, which may be achieved by synthesizing artificial antibodies by means of molecular imprinting techniques. These should also be coupled to simple and low cost devices, such as those of the potentiometric kind, one approach that has been proven successful6. Potentiometric sensors offer the advantage of selectivity and portability for use in point-of-care and have been widely recognized as potential analytical tools in this field. The inherent method is simple, precise, accurate and inexpensive regarding reagent consumption and equipment involved. Thus, this work proposes a new plastic antibody for PSA, designed over the surface of graphene layers extracted from graphite. Charged monomers were used to enable an oriented tailoring of the PSA rebinding sites. Uncharged monomers were used as control. These materials were used as ionophores in conventional solid-contact graphite electrodes. The obtained results showed that the imprinted materials displayed a selective response to PSA. The electrodes with charged monomers showed a more stable and sensitive response, with an average slope of -44.2 mV/decade and a detection limit of 5.8X10-11 mol/L (2 ng/mL). The corresponding non-imprinted sensors showed smaller sensitivity, with average slopes of -24.8 mV/decade. The best sensors were successfully applied to the analysis of serum samples, with percentage recoveries of 106.5% and relatives errors of 6.5%

Novel Prostate Specific Antigen plastic antibody designed withcharged binding sites for an improved protein binding and itsapplication in a biosensor of potentiometric transduction

This work shows that the synthesis of protein plastic antibodies tailored with selected charged monomersaround the binding site enhances protein binding. These charged receptor sites are placed over a neutralpolymeric matrix, thus inducing a suitable orientation the protein reception to its site. This is confirmed bypreparing control materials with neutral monomers and also with non-imprinted template. This concepthas been applied here to Prostate Specific Antigen (PSA), the protein of choice for screening prostate can-cer throughout the population, with serum levels >10 ng/mL pointing out a high probability of associatedcancer.Protein Imprinted Materials with charged binding sites (C/PIM) have been produced by surfaceimprinting over graphene layers to which the protein was first covalently attached. Vinylben-zyl(trimethylammonium chloride) and vinyl benzoate were introduced as charged monomers labellingthe binding site and were allowed to self-organize around the protein. The subsequent polymerizationwas made by radical polymerization of vinylbenzene. Neutral PIM (N/PIM) prepared without orientedcharges and non imprinted materials (NIM) obtained without template were used as controls.These materials were used to develop simple and inexpensive potentiometric sensor for PSA. Theywere included as ionophores in plasticized PVC membranes, and tested over electrodes of solid or liq-uid conductive contacts, made of conductive carbon over a syringe or of inner reference solution overmicropipette tips. The electrodes with charged monomers showed a more stable and sensitive response,with an average slope of -44.2 mV/decade and a detection limit of 5.8 × 10−11mol/L (2 ng/mL). The cor-responding non-imprinted sensors showed lower sensitivity, with average slopes of -24.8 mV/decade.The best sensors were successfully applied to the analysis of serum, with recoveries ranging from 96.9to 106.1% and relative errors of 6.8%

Sarcosine oxidase composite screen-printed electrode for sarcosine determination in biological samples

XIX Meeting of the Portuguese Electrochemical Society - XVI Iberic Meeting of ElectrochemistryProstate Cancer (PCa) is the most common form of cancer in men in Europe with a 61.4 % incidence among all cancer cases and a 12.1 % mortality [1] and, therefore, its early detection is fundamental for increasing the survival rate. Currently, diagnosis and management of patients with PCa is only based on the determination of the biomarker Prostate Specific Antigen (PSA). However, the method used for PCa detection has poor sensitivity and specificity, leading to false negative and false positive test results and many patients are sent to unnecessary biopsy procedures [2]. Therefore, there is a need to seek for new biomarkers and more effective screening. In this work, a biosensor device was developed for the quantification of sarcosine via electrochemical detection of H2O2 (at 0.6 V) generated from the catalyzed oxidation of sarcosine. The detection was carried out after the modification of carbon screen printed electrodes (SPEs) by immobilization of sarcosine oxidase (SOX) on the electrode surface. The strategies used herein included the activation of the carbon films by an electrochemical step and the formation of an NHS/EDAC layer to bond the enzyme to the electrode, the use of metallic or semiconductor nanoparticles layer previously or during the enzyme immobilization. In order to improve the sensor stability and selectivity a polymeric layer with extra enzyme content was further added. The proposed methodology for the detection of sarcosine allowed obtaining a limit of detection (LOD) of 1.6x10-5 mM, using a linear concentration range between 1x10-5 and 1x10-4 mM. The biosensor was successfully applied to the analysis of sarcosine in urine samples.

Lipidomic approach of the biological activity of tacrine and two tacrine-analogues on non-synaptic rat brain mitochondria

Foundation for Science and Technology (FCT), FEDER and COMPET

Chemical characterization, cytotoxic evaluation and anti-SARS-CoV2 activity of plant extracts rich in hydrolysable tannins

info:eu-repo/semantics/publishedVersio

Are olive pomace powders a safe source of bioactives and nutrients?

"First published: 10 September 2020"BACKGROUND Olive oil industry generates significant amounts of semi-solid wastes, namely the olive pomace. Olive pomace is a by-product rich in high-value compounds (e.g. dietary fibre, unsaturated fatty acids, polyphenols) widely explored to obtain new food ingredients. However, conventional extraction methods frequently use organic solvents, while novel eco-friendly techniques have high operational costs. The development of powdered products without any extraction step has been proposed as a more feasible and sustainable approach. RESULTS The present study fractionated and valorised the liquid and pulp fraction of olive pomace obtaining two stable and safe powdered ingredients, namely a liquid-enriched powder (LOPP) and a pulp-enriched powder (POPP). These powders were characterized chemically, and their bioactivity was assessed. LOPP exhibited a significant amount of mannitol (141 g/ kg), potassium (54 g/ kg) and hydroxytyrosol/ derivatives (5 mg/g). POPP exhibited high amount of dietary fibre (620 g/ kg) associated to significant amount of bound phenolics (7.41 mg GAE/ g fibre DW) with substantial antioxidant activity. POPP also contained an unsaturated fatty acids composition similar to olive oil (76\% of total fatty acids) and showed potential as a reasonable source of protein (12 \%). Their functional properties (solubility, water-holding and oil-holding capacity), antioxidant capacity and antimicrobial activity were also assessed, and their biological safety was verified. CONCLUSION The development of olive pomace powders to apply in the food industry could be a suitable strategy to add-value to olive pomace and obtain safe multifunctional ingredients with higher health-promoting effects than dietary fibre and polyphenols itself. This article is protected by copyright. All rights reserved.TBR thanks the Fundação para a Ciência e Tecnologia (FCT), Portugal for PhD grant SFRH/BDE/108271/2015 and the financial support of Association BLC3 – Technology and Innovation Campus. This work was supported by National Funds from FCT – Fundação para a Ciência e a Tecnologia through the project MULTIBIOREFINERY – SAICTPAC/0040/2015 (POCI-01-0145-FEDER-016403). We are also grateful for the scientific collaboration under the FCT project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

Severe and Enduring' Stage in Anorexia Nervosa: Comparing Eating Attitudes, Impairment and Associated Psychopathology

This study aimed to assess differences in eating attitudes, impairment, and related psychopathology at treatment presentation for patients with “Non-severe and enduring Anorexia Nervosa” (illness duration of <7 years) and patients with “severe and enduring Anorexia Nervosa” (illness duration of 7 years or more). One hundred and thirty-nine patients diagnosed with Anorexia Nervosa participated in this study. Participants were interviewed with the Eating Disorder Examination (EDE) and asked to complete several questionnaires at the end of the first treatment appointment. We also explored differences at treatment presentation by considering alternative criteria to define groups, namely a composite of illness duration and clinical impairment (≥16 CIA total score). No differences were found when comparing participants based on illness duration. However, when participants were classified into a different classification scheme: “Non-severe and enduring Anorexia Nervosa” (illness duration <7 years and a CIA total score <16) vs. “severe and enduring Anorexia Nervosa” (illness duration ≥7 years and CIA total score ≥16), significant differences were found in terms of eating pathology, depressive symptomatology, psychological distress, and emotion dysregulation. Further research is needed to better understand the role of illness duration and clinical impairment in informing the course of AN

Corrigendum: Severe and enduring' stage in anorexia nervosa: comparing eating attitudes, impairment and associated psychopathology

In the published article, there was an error in the Funding statement. The funding statement for Eva Conceição was incorrectly displayed as “IF/01219/2014”. The correct Funding statement appears below.FCT - Fundação para a Ciência e a Tecnologia(2020.01538