Translocating the blood-brain barrier using electrostatics

Copyright © 2012 Ribeiro,Domingues, Freire,Santos and Castanho. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.Mammalian cell membranes regulate homeostasis, protein activity, and cell signaling. The charge at the membrane surface has been correlated with these key events. Although mammalian cells are known to be slightly anionic, quantitative information on the membrane charge and the importance of electrostatic interactions in pharmacokinetics and pharmacodynamics remain elusive. Recently, we reported for the first time that brain endothelial cells (EC) are more negatively charged than human umbilical cord cells, using zeta-potential measurements by dynamic light scattering. Here, we hypothesize that anionicity is a key feature of the blood-brain barrier (BBB) and contributes to select which compounds cross into the brain. For the sake of comparison, we also studied the membrane surface charge of blood components—red blood cells (RBC), platelets, and peripheral blood mononuclear cells (PBMC).To further quantitatively correlate the negative zeta-potential values with membrane charge density, model membranes with different percentages of anionic lipids were also evaluated. From all the cells tested, brain cell membranes are the most anionic and those having their lipids mostly exposed, which explains why lipophilic cationic compounds are more prone to cross the blood-brain barrier.Fundação para a Ciência e Tecnologia — Ministério da Educação e Ciência (FCT-MEC, Portugal) is acknowledged for funding (including fellowships SFRH/BD/42158/2007 to Marta M.B. Ribeiro, SFRH/BD/41750/2007 to Marco M. Domingues and SFRH/BD/70423/2010 to João M. Freire) and project PTDC/QUI-BIQ/119509/2010. Marie Curie Industry-Academia Partnerships and Pathways (European Commission) is also acknowledged for funding (FP7-PEOPLE-2007-3-1-IAPP, Project 230654)

Using zeta-potential measurements to quantify peptide partition to lipid membranes

© The Author(s) 2011. This article is published with open access at Springerlink.com.Open Access: This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.Many cellular phenomena occur on the biomembranes. There are plenty of molecules (natural or xenobiotics) that interact directly or partially with the cell membrane. Biomolecules, such as several peptides (e.g., antimicrobial peptides) and proteins, exert their effects at the cell membrane level. This feature makes necessary investigating their interactions with lipids to clarify their mechanisms of action and side effects necessary. The determination of molecular lipid/water partition constants (Kp) is frequently used to quantify the extension of the interaction. The determination of this parameter has been achieved by using different methodologies, such as UV-Vis absorption spectrophotometry, fluorescence spectroscopy and ζ-potential measurements. In this work, we derived and tested a mathematical model to determine the Kp from ζ-potential data. The values obtained with this method were compared with those obtained by fluorescence spectroscopy, which is a regular technique used to quantify the interaction of intrinsically fluorescent peptides with selected biomembrane model systems. Two antimicrobial peptides (BP100 and pepR) were evaluated by this new method. The results obtained by this new methodology show that ζ-potential is a powerful technique to quantify peptide/lipid interactions of a wide variety of charged molecules, overcoming some of the limitations inherent to other techniques, such as the need for fluorescent labeling.This work was partially supported by project PTDC/QUI/ 69937/2006 from Fundação para a Ciência e Tecnologia-Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES, Portugal), and by Fundação Calouste Gulbenkian (Portugal). JMF and MMD also thank FCT-MCTES for grants IMM/BT/37-2010 and SFRH/BD/41750/2007, respectively

Controlo mecânico de infestantes

As infestantes são plantas indesejáveis que crescem juntamente com as plantas cultivadas e que interferem no seu desenvolvimento normal. As infestantes podem ser uma das principais causas da diminuição do rendimento das culturas, porque competem com elas para o espaço, para a água, luz solar, nutrientes e dióxido de carbono, podem segregar substâncias alelopáticas, ser o meio no qual temporariamente se instalam alguns organismos responsáveis por inúmeras pragas e doenças que atacam as culturas dificultando assim o combate às mesmas, dificultam a colheita quer esta seja manual ou mecanizada, podem contaminar o produto final, depreciando-o e, asseguram a reinfestação para as culturas seguintes. O controlo de infestantes ter-se-á iniciado quando o homem deixou a de ser nómada e de assegurar as suas necessidades através da colheita de frutos e da caça e passou após a “domesticação“ das espécies animais e vegetais a fazer agricultura, tornando-se sedentário. Portanto, desde o início da agricultura, que o homem tem feito grandes esforços para controlar as plantas infestantes, primeiro à mão, depois com o uso de alguns artefactos, ferramentas e equipamentos para melhorar a eficiência no seu controlo. Hoje existem equipamentos mecânicos sofisticados tal como, substâncias químicas ou biológicas que permitem o seu controlo prevenindo ou retardando a sua germinação ou crescimento. Interferência das plantas infestantes com a cultura pode gerar perdas significativas, na qualidade e quantidade de alimentos produzidos, desperdiçando enormes quantidades de energia, especialmente não renováveis. Os custos no controlo e os efeitos sobre os rendimentos são muito variáveis, dependendo do agricultor, das espécies de plantas infestantes e da estratégia ou estratégias adoptadas para garantir a eficácia no controlo. Nas últimas cinco décadas têm vindo a fazer-se significativos avanços científicos e tecnológicos na criação de estratégias para o aumento da eficácia no controlo de infestantes seja mecanicamente, seja através da utilização de substâncias químicas ou biológicas menos tóxicas para o homem, menos agressivas ao meio ambiente, com menores custos de produção e ao mesmo tempo, mais selectivas para as culturas onde são aplicadas. A alternativa ao controlo químico de infestantes através da aplicação de herbicidas é o controlo mecânico pela utilização de diversas alfaias agrícolas, tais como a charrua de aivecas, a charrua de discos, o escarificador de braços rígidos, o escarificador de braços flexíveis (vibrocultor) e a fresa. O controlo mecânico de infestantes poderá ser levado a cabo também por máquinas de corte, como por exemplo, as gadanheiras. Cortar as infestantes numa fase de desenvolvimento antes da produção de semente evita a sua propagação. Se o agricultor optar pela sementeira directa como técnica de instalação das culturas, a única alternativa que tem para o controlo de infestantes é a química, mas se optar pelo sistema de mobilização tradicional ou pela mobilização reduzida poderá controlar as infestantes, química e/ou mecanicamente. A eficácia das diferentes alfaias no controlo de infestantes depende da própria alfaia, da época do ano em que se realiza esse controlo, do estado do solo, das espécies de infestantes presentes e seu estádio de desenvolvimento. Iremos no presente trabalho, referir os aspectos mais importantes do controlo mecânico de infestantes

Blending In or Standing Out? Gendered Political Communication in 24 Democracies

Women in male-dominated organizations often must adopt more stereotypical masculine traits to advance within those hierarchies. While politics, historically male-dominated, should induce women to blend in, increasing numbers of women in parliaments may give women the opportunity to stand out by not adopting a masculine style. This paper investigates how these contradictory incentives influence female Members of Parliament (MPs) in 24 democracies between 1987 and 2022, applying machine learning to 6.8 million parliamentary speeches to measure how feminine is their speaking style. Findings indicate a socialization effect, whereby women adopt a more masculine style the longer they stay in office, even after controlling for their speeches’ topics. The effect is strongest for women in socially progressive parties. This research highlights the role of parliaments as gendered workplaces, which still lead women to adapt to the male norm, and helps us understand the incentives that shape how women represent women in parliament.Women rising in male-dominated environments Members of Parliament and speech data Measuring gendered language Explaining gendered language Conclusion Acknowledgments Supporting information Reference

Pharmaceutical innovations: the grand challenges ahead

Copyright © 2020 Aroeira and Castanho. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Lifestyles are evolving rapidly due to swiftly evolving technologies rapid exchange of information across the globe, and facilitated mobility of people across long distances. In addition, climate changes accelerate the geographical dynamics of disease. The result is that both communicable and non-communicable diseases pose challenges never faced before and the perception of the way pharmaceutical sciences are dealing with such changes is under unprecedented scrutiny. The discredit in science-based solutions has a tremendous societal impact and is detrimental to evidence-based pharmacology at large. Pharmaceutical innovation that target the needs of healthy living and meet the expectation of society are urgently needed and are a worthy effect of both industrial and academic researchers. A reflection on the grand challenges ahead in thus timely and appropriate.This work was supported by La Caixa Foundation (grant reference: IMM/BPD/107-2018 to RA).info:eu-repo/semantics/publishedVersio

The mechanism of action of antimicrobial peptides : lipid vesicles vs. bacteria

Copyright © 2012 Melo and Castanho. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.The authors acknowledge the European FP7-PEOPLE-2009-IEF-254559 grant to Manuel N. Melo and the Fundação para a Ciência e a Tecnologia (Portugal) project PTDC/QUI-BIQ/112929/2009

Anticancer peptides : prospective innovation in cancer therapy

© Springer International Publishing Switzerland 2016Current cancer treatments require improvements in selectivity and efficacy. Surgery, radiation, and chemotherapy approaches result in patient’s suffering over time due to the development of severe side-effects that simultaneously condition adherence to therapy. Biologically active peptides, in particular antimicrobial peptides (AMPs), are versatile molecules in terms of biological activities. The cytotoxic activities of several AMPs turn this group of molecules into an amazing pool of new templates for anticancer drug development. However, several unmet challenges limit application of peptides in cancer therapy. The mechanism(s) of action of the peptides need better description and understanding, and innovative targets have to be discovered and explored, facilitating drug design and development. In this chapter, we explore the natural occurring AMPs as potential new anticancer peptides (ACPs) for cancer prevention and treatment. Their modes of action, selectivity to tumor compared to normal cells, preferential targets, and applications, but also their weaknesses, are described and discussed.info:eu-repo/semantics/publishedVersio

Resazurin reduction-based assays revisited: guidelines for accurate reporting of relative differences on metabolic status

© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Cell viability and metabolic activity are ubiquitous parameters used in biochemistry, molecular biology, and biotechnological studies. Virtually all toxicology and pharmacological projects include at some point the evaluation of cell viability and/or metabolic activity. Among the methods used to address cell metabolic activity, resazurin reduction is probably the most common. At variance with resazurin, resorufin is intrinsically fluorescent, which simplifies its detection. Resazurin conversion to resorufin in the presence of cells is used as a reporter of metabolic activity of cells and can be detected by a simple fluorometric assay. UV–Vis absorbance is an alternative technique but is not as sensitive. In contrast to its wide empirical “black box” use, the chemical and cell biology fundamentals of the resazurin assay are underexplored. Resorufin is further converted to other species, which jeopardizes the linearity of the assays, and the interference of extracellular processes has to be accounted for when quantitative bioassays are aimed at. In this work, we revisit the fundamentals of metabolic activity assays based on the reduction of resazurin. Deviation to linearity both in calibration and kinetics, as well as the existence of competing reactions for resazurin and resorufin and their impact on the outcome of the assay, are addressed. In brief, fluorometric ratio assays using low resazurin concentrations obtained from data collected at short time intervals are proposed to ensure reliable conclusions.The project leading to these results has received funding from “la Caixa” Foundation and FCT, I.P. under the project code [LCF/PR/HR21/00605], BREAST-BRAIN-N-BBB.info:eu-repo/semantics/publishedVersio

The use of antibody-antibiotic conjugates to fight bacterial infections

Copyright © 2022 Cavaco, Castanho and Neves. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.The emergence of antimicrobial resistance (AMR) is rapidly increasing and it is one of the significant twenty-first century’s healthcare challenges. Unfortunately, the development of effective antimicrobial agents is a much slower and complex process compared to the spread of AMR. Consequently, the current options in the treatment of AMR are limited. One of the main alternatives to conventional antibiotics is the use of antibodyantibiotic conjugates (AACs). These innovative bioengineered agents take advantage of the selectivity, favorable pharmacokinetic (PK), and safety of antibodies, allowing the administration of more potent antibiotics with less off-target effects. Although AACs’ development is challenging due to the complexity of the three components, namely, the antibody, the antibiotic, and the linker, some successful examples are currently under clinical studies.The project leading to these results has received funding from “la Caixa” Foundation (ID 100010434), under the agreement LCF/PR/HR17/52150011 and from Portuguese Funding Agency, Fundação para a Ciência e Tecnologia (FCT IP, grants PD/BD/128281/2017 and DL 7/2016/CP1451/CT0023).info:eu-repo/semantics/publishedVersio

From antimicrobial to anticancer peptides : a review

Antimicrobial peptides (AMPs) are part of the innate immune defense mechanism of many organisms. Although AMPs have been essentially studied and developed as potential alternatives for fighting infectious diseases, their use as anticancer peptides (ACPs) in cancer therapy either alone or in combination with other conventional drugs has been regarded as a therapeutic strategy to explore. As human cancer remains a cause of high morbidity and mortality worldwide, an urgent need of new, selective, and more efficient drugs is evident. Even though ACPs are expected to be selective toward tumor cells without impairing the normal body physiological functions, the development of a selective ACP has been a challenge. It is not yet possible to predict antitumor activity based on ACPs structures. ACPs are unique molecules when compared to the actual chemotherapeutic arsenal available for cancer treatment and display a variety of modes of action which in some types of cancer seem to co-exist. Regardless the debate surrounding the definition of structure-activity relationships for ACPs, great effort has been invested in ACP design and the challenge of improving effective killing of tumor cells remains. As detailed studies on ACPs mechanisms of action are crucial for optimizing drug development, in this review we provide an overview of the literature concerning peptides' structure, modes of action, selectivity, and efficacy and also summarize some of the many ACPs studied and/or developed for targeting different solid and hematologic malignancies with special emphasis on the first group. Strategies described for drug development and for increasing peptide selectivity toward specific cells while reducing toxicity are also discussed.The authors thank Fundação para a Ciência e a Tecnologia (FCT- MEC, Portugal) for funding—PTDC/QUI-BIO/112929/2009. Diana Gaspar also acknowledges FCT for fellowship SFRH/BPD/ 73500/2010 and A. Salome Veiga for funding within the FCT Investigator Programme (IF/00803/2012