Secretion of the fungal toxin candidalysin is dependent on conserved precursor peptide sequences.

The opportunistic fungal pathogen Candida albicans damages host cells via its peptide toxin, candidalysin. Before secretion, candidalysin is embedded in a precursor protein, Ece1, which consists of a signal peptide, the precursor of candidalysin and seven non-candidalysin Ece1 peptides (NCEPs), and is found to be conserved in clinical isolates. Here we show that the Ece1 polyprotein does not resemble the usual precursor structure of peptide toxins. C. albicans cells are not susceptible to their own toxin, and single NCEPs adjacent to candidalysin are sufficient to prevent host cell toxicity. Using a series of Ece1 mutants, mass spectrometry and anti-candidalysin nanobodies, we show that NCEPs play a role in intracellular Ece1 folding and candidalysin secretion. Removal of single NCEPs or modifications of peptide sequences cause an unfolded protein response (UPR), which in turn inhibits hypha formation and pathogenicity in vitro. Our data indicate that the Ece1 precursor is not required to block premature pore-forming toxicity, but rather to prevent intracellular auto-aggregation of candidalysin sequences.

Ecological Niche-Inspired Genome Mining Leads to the Discovery of Crop-Protecting Nonribosomal Lipopeptides Featuring a Transient Amino Acid Building Block.

Investigating the ecological context of microbial predator-prey interactions enables the identification of microorganisms, which produce multiple secondary metabolites to evade predation or to kill the predator. In addition, genome mining combined with molecular biology methods can be used to identify further biosynthetic gene clusters that yield new antimicrobials to fight the antimicrobial crisis. In contrast, classical screening-based approaches have limitations since they do not aim to unlock the entire biosynthetic potential of a given organism. Here, we describe the genomics-based identification of keanumycins A-C. These nonribosomal peptides enable bacteria of the genus Pseudomonas to evade amoebal predation. While being amoebicidal at a nanomolar level, these compounds also exhibit a strong antimycotic activity in particular against the devastating plant pathogen Botrytis cinerea and they drastically inhibit the infection of Hydrangea macrophylla leaves using only supernatants of Pseudomonas cultures. The structures of the keanumycins were fully elucidated through a combination of nuclear magnetic resonance, tandem mass spectrometry, and degradation experiments revealing an unprecedented terminal imine motif in keanumycin C extending the family of nonribosomal amino acids by a highly reactive building block. In addition, chemical synthesis unveiled the absolute configuration of the unusual dihydroxylated fatty acid of keanumycin A, which has not yet been reported for this lipodepsipeptide class. Finally, a detailed genome-wide microarray analysis of Candida albicans exposed to keanumycin A shed light on the mode-of-action of this potential natural product lead, which will aid the development of new pharmaceutical and agrochemical antifungals.

Candidalysin Is the Hemolytic Factor of Candida albicans.

Candida albicans produces an important virulence factor, the hypha-associated Ece1-derived secreted peptide toxin candidalysin, which is crucial for the establishment of mucosal and systemic infections. C. albicans has also long been known to be hemolytic, yet the hemolytic factor has not been clearly identified. Here, we show that candidalysin is the hemolytic factor of C. albicans. Its hemolytic activity is modulated by fragments of another Ece1 peptide, P7. Hemolysis by candidalysin can be neutralized by the purinergic receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). PPADS also affects candidalysin's ability to intercalate into synthetic membranes. We also describe the neutralization potential of two anti-candidalysin nanobodies, which are promising candidates for future anti-Candida therapy. This work provides evidence that the historically proposed hemolytic factor of C. albicans is in fact candidalysin and sheds more light on the complex roles of this toxin in C. albicans biology and pathogenicity.

Fungal factors involved in host immune evasion, modulation and exploitation during infection.

Human and plant pathogenic fungi have a major impact on public health and agriculture. Although these fungi infect very diverse hosts and are often highly adapted to specific host niches, they share surprisingly similar mechanisms that mediate immune evasion, modulation of distinct host targets and exploitation of host nutrients, highlighting that successful strategies have evolved independently among diverse fungal pathogens. These attributes are facilitated by an arsenal of fungal factors. However, not a single molecule, but rather the combined effects of several factors enable these pathogens to establish infection. In this review, we discuss the principles of human and plant fungal pathogenicity mechanisms and discuss recent discoveries made in this field.

Interprofessional collaboration from nurses and physicians ­ A triangulation of quantitative and qualitative data / Interprofessionelle Zusammenarbeit Pflegefachpersonen und Ärzteschaft. Eine Triangulation quantitativer und qualitativer Daten

Background: Interprofessional collaboration between nurses and physicians is a recurrent challenge in daily clinical practice. To ameliorate the situation, quantitative or qualitative studies are conducted. However, the results of these studies have often been limited by the methods chosen. Aim: To describe the synthesis of interprofessional collaboration from the nursing perspective by triangulating quantitative and qualitative data. Method: Data triangulation was performed as a sub-project of the interprofessional Sinergia DRG Research program. Initially, quantitative and qualitative data were analyzed separately in a mixed methods design. By means of triangulation a "meta-matrix" resulted in a four-step process. Results: The "meta-matrix" displays all relevant quantitative and qualitative results as well as their interrelations on one page. Relevance, influencing factors as well as consequences of interprofessional collaboration for patients, relatives and systems become visible. Conclusion: For the first time, the interprofessional collaboration from the nursing perspective at five Swiss hospitals is shown in a "meta-matrix". The consequences of insufficient collaboration between nurses and physicians are considerable. This is why it's necessary to invest in interprofessional concepts. In the "meta-matrix" the factors which influence the interprofessional collaboration positively or negatively are visible.

Negociando saberes e poderes: a Política Nacional de Atenção Integral à Saúde do Homem e a Sociedade Brasileira de Urologia / Negotiating knowledge and power: the National Policy for Comprehensive Men’ s Healthcare and the Brazilian Society of Urology

Resumo O objetivo do artigo é evidenciar as negociações e disputas entre saberes e poderes na história da Política Nacional de Atenção Integral à Saúde do Homem, a partir da criação da Área Técnica de Saúde do Homem, instituída em 2008 no Departamento de Ações Programáticas Estratégicas no Ministério da Saúde. Acompanhamos a posição da Sociedade Brasileira de Urologia no processo de constituição da política. Recuperamos a disputa colocada em discurso tendo como referência as entrevistas realizadas com gestores do Ministério da Saúde e com o representante auxiliar do Fundo de População das Nações Unidas no Brasil. Nas aparições do sujeito de direitos à saúde, analisamos a visibilidade do corpo masculino nos signos das resistências às intervenções do biopoder.

Abstract The aim of this article is to demonstrate the negotiations and disputes between different kinds of knowledge and power in the history of the Brazilian National Policy for Comprehensive Men’s Healthcare based on the creation in 2008 of the Technical Area for Men’s Health within the Ministry of Health’s Department of Strategic Programs. We observed the Brazilian Society of Urology’s position as the policy was being drawn up, including the discourse adopted, based on interviews held with managers from the Ministry of Health and the assistant representative of the United Nations Population Fund in Brazil. We analyzed the visibility of the male body in signs of resistance to the interventions of biopower in the expression of the right to health.

[Negotiating knowledge and power: the National Policy for Comprehensive Men's Healthcare and the Brazilian Society of Urology]. / Negociando saberes e poderes: a Política Nacional de Atenção Integral à Saúde do Homem e a Sociedade Brasileira de Urologia.

The aim of this article is to demonstrate the negotiations and disputes between different kinds of knowledge and power in the history of the Brazilian National Policy for Comprehensive Men's Healthcare based on the creation in 2008 of the Technical Area for Men's Health within the Ministry of Health's Department of Strategic Programs. We observed the Brazilian Society of Urology's position as the policy was being drawn up, including the discourse adopted, based on interviews held with managers from the Ministry of Health and the assistant representative of the United Nations Population Fund in Brazil. We analyzed the visibility of the male body in signs of resistance to the interventions of biopower in the expression of the right to health.

A política nacional de atenção integral à saúde do homem e suas engrenagens biopolíticas: o uso do conceito de gênero como regime de luzes / The national policy for integral attention to men's health and its biopolitics gear: the use of the concept of gender as a light regime

O artigo analisa a Política Nacional de Atenção Integral à Saúde do Homem do governo brasileiro. Foram analisados os documentos "Princípios e diretrizes da Política Nacional de Atenção Integral à Saúde do Homem", "Princípios e diretrizes da Política Nacional de Atenção Integral à Saúde da Mulher"; e entrevistas realizadas com gestores federais da Área Técnica de Saúde do Homem. A análise do discurso a partir de Michel Foucault permitiu lidar com o corpus em suas condições de produção mediante um gesto analítico-descritivo de manejo dos enunciados dispersos. O argumento central é o de que a Política de saúde tem um "gênero" como uma rede discursiva que produz os sujeitos referências de sua inteligibilidade. Destacam-se na análise a emergência do sujeito feminino do cuidado, representada pela mulher-mãe higiênica e mulher-sujeito; e do homem como sujeito de direitos à saúde sob o signo de acontecimento de seu nascimento

The article examines the National Policy of Comprehensive Men and Women's Healthcare of the Brazilian government. Documents analyzed were: "Principles and guidelines of the National Policy of Comprehensive Men's Healthcare", "Principles and guidelines of the National Policy of Comprehensive Women's Healthcare", and interviews to federal managers of the Technical Field of Men's Healthcare. Discourse analysis from Michel Foucault allowed to deal with the corpus in their production conditions through an analytical -descriptive gesture handling of scattered statements. The central argument is that the health policy analyzed has a "gender" as a discursive network that produces the subject references its intelligibility. Stand - in analyzes the emergence of the female subject of care, represented by the woman-mother hygienic and woman -subject, and the man as the subject of rights to health under the sign of newness of his birth. The dialogue with Judith Butler and Jacques Derrida is one of the enablers axes theoretical analysis of the issues that build the product , in addition to the title itself also seeks to highlight , namely the biopolitics gears that move the policy in question.

A Política Nacional de Atenção Integral à Saúde do Homem e suas engrenagens biopolíticas: o uso do conceito de gênero como regime de luzes / The National Policy for Integral Attention to Men’s Health and its biopolitics gear: the use of the concept of gender as a light regime

O artigo analisa a Política Nacional de Atenção Integral à Saúde do Homem do governo brasileiro. Foram analisados os documentos “Princípios e diretrizes da Política Nacional de Atenção Integral à Saúde do Homem”, “Princípios e diretrizes da Política Nacional de Atenção Integral à Saúde da Mulher”; e entrevistas realizadas com gestores federais da Área Técnica de Saúde do Homem. A análise do discurso a partir de Michel Foucault permitiu lidar com o corpus em suas condições de produção mediante um gesto analítico-descritivo de manejo dos enunciados dispersos. O argumento central é o de que a Política de saúde tem um “gênero” como uma rede discursiva que produz os sujeitos referências de sua inteligibilidade. Destacam-se na análise a emergência do sujeito feminino do cuidado, representada pela mulher-mãe higiênica e mulher-sujeito; e do homem como sujeito de direitos à saúde sob o signo de acontecimento de seu nascimento.(AU)

The article examines the National Policy of Comprehensive Men and Women’s Healthcare of the Brazilian government. Documents analyzed were: “Principles and guidelines of the National Policy of Comprehensive Men’s Healthcare”, “Principles and guidelines of the National Policy of Comprehensive Women’s Healthcare”, and interviews to federal managers of the Technical Field of Men’s Healthcare. Discourse analysis from Michel Foucault allowed to deal with the corpus in their production conditions through an analytical -descriptive gesture handling of scattered statements. The central argument is that the health policy analyzed has a “gender” as a discursive network that produces the subject references its intelligibility. Stand - in analyzes the emergence of the female subject of care, represented by the woman-mother hygienic and woman -subject, and the man as the subject of rights to health under the sign of newness of his birth. The dialogue with Judith Butler and Jacques Derrida is one of the enablers axes theoretical analysis of the issues that build the product , in addition to the title itself also seeks to highlight , namely the biopolitics gears that move the policy in question .(AU)

Predicting Activators and Inhibitors of the Breast Cancer Resistance Protein (ABCG2) and P-Glycoprotein (ABCB1) Based on Mechanistic Considerations.

Colocalized in membrane barriers, the ABC transporters ABCB1 and ABCG2 strongly contribute to multidrug resistance (MDR). Here we investigate the as yet unknown mechanisms of activation and inhibition of ABCG2. For this purpose we measured the ATPase activity of ABCG2 and ABCB1 as a function of allocrite concentration using a calibration set of 30 diverse compounds and a validation set of 23 compounds. We demonstrate that ABCG2 is activated at low and inhibited at high allocrite concentrations, yielding bell-shaped activity curves. With an ATP regeneration assay we prove that the inhibitory part is indeed due to a decrease in activity because of high allocrite load in the transporter. However, inhibition is only observed if the membrane solubility of allocrites is sufficiently high. The concentrations of half-maximum activation and inhibition are at least 10-fold lower for ABCG2 than for ABCB1. Because ABCG2 binds its allocrites with higher affinity than ABCB1, it can extract hydrophilic, nonamphiphilic, and highly charged compounds out of the lipid membrane, typically exhibiting low lipid-water partition coefficients, but is inhibited by hydrophobic, amphiphilic, and moderately charged compounds, with high lipid-water partition coefficients. In contrast, ABCB1 is barely interacting with hydrophilic compounds, but is activated by hydrophobic compounds. We show that hydrophobicity, amphiphilicity, and charge have a dual role; they predict, on the one hand, allocrites' lipid-water partition coefficient and, on the other hand, the transporters' preference for the chemical nature of allocrites. Parameters reflecting hydrophobicity, amphiphilicity, and charge are therefore sufficient for differentiating between allocrites, activators, and inhibitors of ABCB1 and ABCG2.

Allocrite Sensing and Binding by the Breast Cancer Resistance Protein (ABCG2) and P-Glycoprotein (ABCB1).

The ATP binding cassette (ABC) transporters ABCG2 and ABCB1 perform ATP hydrolysis-dependent efflux of structurally highly diverse compounds, collectively called allocrites. Whereas much is known about allocrite-ABCB1 interactions, the chemical nature and strength of ABCG2-allocrite interactions have not yet been assessed. We quantified and characterized interactions of allocrite with ABCG2 and ABCB1 using a set of 39 diverse compounds. We also investigated potential allocrite binding sites based on available transporter structures and structural models. We demonstrate that ABCG2 binds its allocrites from the lipid membrane, despite their hydrophilicity. Hence, binding of allocrite to both transporters is a two-step process, starting with a lipid-water partitioning step, driven mainly by hydrophobic interactions, followed by a transporter binding step in the lipid membrane. We show that binding of allocrite to both transporters increases with the number of hydrogen bond acceptors in allocrites. Scrutinizing the transporter translocation pathways revealed ample hydrogen bond donors for allocrite binding. Importantly, the hydrogen bond donor strength is, on average, higher in ABCG2 than in ABCB1, which explains the higher measured affinity of allocrite for ABCG2. π-π stacking and π-cation interactions play additional roles in binding of allocrite to ABCG2 and ABCB1. With this analysis, we demonstrate that these membrane-mediated weak electrostatic interactions between transporters and allocrites allow for transporter promiscuity toward allocrites. The different sensitivities of the transporters to allocrites' charge and amphiphilicity provide transporter specificity. In addition, we show that the different hydrogen bond donor strengths in the two transporters allow for affinity tuning.

Masculinidades e práticas de saúde: retratos da experiência de pesquisa em Florianópolis/SC / Masculinities and health practices: portraits of a research experience in Florianopolis (SC), Brazil

Este artigo tem o objetivo de apresentar parte dos resultados da pesquisa multicêntrica "Masculinidades e Práticas de Saúde", em que foram investigados, junto à população masculina de Florianópolis (SC), os significados sobre saúde/doença e práticas de cuidados com a saúde por ela desenvolvidas. Para tal, foram realizados sete grupos focais com homens usuários dos serviços de saúde, entrevistas com os profissionais que compõem a equipe técnica de saúde sexual e reprodutiva do Hospital Universitário (HU) da Universidade Federal de Santa Catarina (UFSC), bem como 260 entrevistas com homens. Investigou também, junto à equipe de profissionais do serviço (médicos, enfermeiros, psicólogos, assistentes sociais, gestores em saúde), as discursividades de gênero em relação aos cuidados com a saúde e promoção do acesso de homens aos serviços. A proposta filosófico-metodológica de Judith Butler sobre gênero inteligível, performatividade e suas implicações na constituição de sujeitos em saúde foi o vetor de análise do material empírico da pesquisa, sem perder de vista o projeto biopolítico em que estes enunciados são possíveis de serem formulados, principalmente no que se refere ao SUS e a seus princípios de universalidade, integralidade e equidade. A categoria gênero é elemento de grande importância no padrão dos riscos de saúde nos homens e na forma como estes percebem, usam e cuidam de seus corpos, tanto como possibilidade de transformação de valores como na reiteração dos vigentes.

This paper aims to present partial results of the multicentered research "Masculinities and Health Practices". This research explored the meanings of health and illness among the male population in Florianopolis and the healthcare adopted by them. The research methodology was based on seven focal groups with male patients of the local health services, interviews with the professionals who take part of the sexual health and reproductive team of the Santa Catarina University Hospital, as well as 260 interviews with males. It also explored gender discursive practices of the medical team (doctors, nurses, psychologists, social assistants, and health professionals in general) regarding healthcare and the availability of the services for the male population. Judith Butler's philosophical and methodological proposals regarding intelligible gender, performances and its consequences for the constitution of the subjects was the theoretical vector of analyses of the empirical material. The paper also considers the biopolitical project that encapsulated these ideas, in particular the Unified Health System (SUS) and its principles of universality, integral care and equity. The concept of gender is a key element of the health risk pattern among the males and how they perceive, act and take care of their bodies, both as possibility of values transformations or reiteration of current values.

[Risk factors for falls and fall-related injuries in an acute care hospital--a retrospective case control study]. / Risikofaktoren für Stürze und sturzbedingte Verletzungen im Akutspital - Eine retrospektive Fall-Kontroll-Studie.

Falls in patient are a major problem in acute care institutions because of direct and indirect consequences. The objectives of the present retrospective case control study were to explore predictors of falls and fall-related injuries in hospitalised patients in a department of internal medicine at a Swiss acute care hospital. The sample included 228 hospitalised patients with a fall and 228 patients without a fall as control subjects, matched by age, gender and medical diagnosis. The cases were further analysed, whereby injured patients were compared with uninjured ones. Data were obtained from patient records and the hospital information system (HIS). Patients with a previous fall, mobility disorders, altered cognition, altered urinary and fecal excretions, intake of sedativa or other psychotropic drugs, a higher amount of comorbidities and case mix, as well as a prolonged length of stay were significantly more frequent in the cases than in the controls. Patients with falls and controls showed no differences in terms of impaired vision, intake of five and more medications, or diuretica. Analysis by logistic regression revealed mobility disorder (OR 5,2; 95 % VI 2.39-11.44) and altered excretions (OR 1,8; 95 % VI 1.02-3.31) as significant main effects and there also were significant interactions between previous falls, intake of sedativa or other psychopharmaca and altered cognition. No difference could be seen in any variable between patients with a fall-related injury and no injury. Therefore a predictive profile could be found for patients at risk to fall which might be helpful for further prevention strategies. However, no predictive marker for fall-related injuries could be defined in the present study.

Kinetic mechanism and catalysis of a native-state prolyl isomerization reaction.

Folding of tendamistat is a rapid two-state process for the majority of the unfolded molecules. In fluorescence-monitored refolding kinetics about 8% of the unfolded molecules fold slowly (lambda=0.083s(-1)), limited by peptidyl-prolyl cis-trans isomerization. This is significantly less than expected from the presence of three trans prolyl-peptide bonds in the native state. In interrupted refolding experiments we detected an additional very slow folding reaction (lambda=0.008s(-1) at pH 2) with an amplitude of about 12%. This reaction is caused by the interconversion of a highly structured intermediate to native tendamistat. The intermediate has essentially native spectroscopic properties and about 2% of it remain populated in equilibrium after folding is complete. Catalysis by human cyclophilin 18 identifies this very slow reaction as a prolyl isomerization reaction. This shows that prolyl-isomerases are able to efficiently catalyze native state isomerization reactions, which allows them to influence biologically important regulatory conformational transitions. Folding kinetics of the proline variants P7A, P9A, P50A and P7A/P9A show that the very slow reaction is due to isomerization of the Glu6-Pro7 and Ala8-Pro9 peptide bonds, which are located in a region that makes strong backbone and side-chain interactions to both beta-sheets. In the P50A variant the very slow isomerization reaction is still present but native state heterogeneity is not observed any more, indicating a long-range destabilizing effect on the alternative native state relative to N. These results enable us to include all prolyl and non-prolyl peptide bond isomerization reactions in the folding mechanism of tendamistat and to characterize the kinetic mechanism and the energetics of a native-state prolyl isomerization reaction.

Brown trout Salmo trutta f. Fario liver ultrastructure as a biomarker for assessment of small stream pollution.

T he effects of environmental pollutants from two small streams in south-west Germany on the liver of brown trout (S almo trutta f. Fario) were studied as biomarkers by means of quantitative and semi-quantitative electron microscopy, and quantitatively by morphometrical measurements. Cellular damage was assessed semi-quantitatively based on a classification of ultrastructural responses. Both methods revealed more severe cellular effects in the liver of trout which had been exposed to the highly polluted stream than in those exposed to the lightly polluted river. Morphometrical studies showed a significant reduction of glycogen storage and a significant increase in number of mitochondria, peroxisomes and cisternae of the endoplasmic reticulum. The biomarker responses of this study were correlated with the results obtained by limnological and analytical investigations, and reflect the levels of pollution in each stream.