Sexual behaviour and HIV/sexually transmitted infection risk behaviours in the general population of Slovenia, a low HIV prevalence country in central Europe.

OBJECTIVES: To describe sexual and HIV/sexually transmitted infection (STI) risk behaviours in Slovenia. METHODS: A nationally representative cross-sectional survey of the general population aged 18-49 years in 1999-2001 was conducted. The data were collected by face-to-face interviews and anonymous self-administered questionnaires. Statistical methods for complex survey data were used. RESULTS: 849 men and 903 women were interviewed. In the past 5 years, both men and women reported a median of one heterosexual partner (means 3.2, 1.5, respectively), concurrent heterosexual partnerships were reported by 24.4% of men and 8.2% of women, heterosexual sex with non-Slovenian partners by 12.6% of men and 12.2% of women, forced sex by 4.8% of women, paid heterosexual sex by 2.6% of men, sex with another man by 0.6% of men and heterosexual sex with an injecting drug user by 1.2% of men and 1.3% of women. In the past year, 22.7% of men and 9.5% of women reported forming at least one new heterosexual partnership. The mean numbers of episodes of heterosexual sex in the previous 4 weeks were 6.1 for men and 6.0 for women. Consistent and inconsistent condom use was reported more frequently among men reporting multiple female partners and those not married or cohabiting. CONCLUSIONS: Recent patterns of reported sexual behaviour are consistent with a low risk of HIV and STI transmission in Slovenia. The results will inform Slovenian sexual health policies including HIV/STI prevention, and are particularly valuable because population-based data on HIV/STI risk behaviour have not previously been available in low HIV prevalence countries of central Europe

Leveraging vectored vaccine candidates manufacturing to GMP compatible bioprocesse

Background Vectored vaccines are very efficient in the in vivo delivery of antigens either in the form of antigen protein and peptides or genetic material. The bioprocess of vectored vaccines poses however several challenge since the viral particles to be effective must maintain their infectivity. Lentiviral and adenoviral vectors are among the particles more used in the treatment of cancer diseases modulating the immune system. Both viral vectors are currently produced in transient upstream process. While the adenoviral vectors are produced at high titers the lentiviral vector upstream process still requires further improvement. The non-lytic nature of lentivirus enables the design of stable cell lines which may improve its yields through perfusion and longer term productions, reducing costs. The application of novel methods for the downstream processing such as continuous purification will contribute to increase the yield and lower the overall cost of the manufacturing processes. Experimental approach At the upstream process, many of the challenges lentiviral bioproducts present in its manufacturing are related to the apoptosis-leading cytotoxicity of some of the vector components. Supported on our long track experience and enabling tools developed for gammaretrovirus manufacturing, we undertook the challenge of establishing a constitutive stable lentiviral producer cell line. To address this challenge we proposed to eliminate or reduce the cytotoxicity of the lentiviral vector expression components. At the downstream process lentiviral vectors face the challenges common to retroviridae family of vectors namely short half-lives at room temperature, sensitivity to pH variations and salt concentrations, and shear stress. The purification strategy developed was designed to be based on disposable and easily scalable technologies. A final concentration achieving 108 TU mL-1 was targeted since the concentration step itself allows to reduce the burden on process and improve the transduction efficiency. To address the high doses requirements we will report an improved oncolytic adenovirus purification process for phase I and II clinical trials and present a case on the use of Polysorb 20 as a replacement for Triton X-100 during cell lysis. Product recovery, potency, purity and the effect of manufacturing holding points will be discussed. Results and discussion A lentiviral producer cell line constitutively producing titers above 106 TU.mL-1.day-1 was established. The cell line showed to be stable, consistently maintaining vector productivity over one month in the absence of antibiotics. At the bioreaction process it was possible to maintain the cells continuously producing over 10 days. At downstream we implemented scalable protocols for lentiviral and adenoviral vectors that is easy to transfer to GMP environment, combining microfiltration, anion-exchange, and ultrafiltration membranes technologies toward maximization of infectious virus recovery, allowing generation of clinical-grade viral vectors without the need for cleaning validation in a cost-effective manner. Herein we will present and discuss the challenges on the biomanufacturing of lentiviral as well as adenoviral virus, the strategies and novel technologies to be adopted in order to enable a faster development of novel vectored vaccine candidates focusing on several case studies, supported by process technology innovation

The carboxylic acid transporters Jen1 and Jen2 affect the architecture and fluconazole susceptibility of Candida albicans biofilm in the presence of lactate

Candida albicans has the ability to adapt to different host niches, often glucose-limited but rich in alternative carbon sources. In these glucose-poor microenvironments, this pathogen expresses JEN1 and JEN2 genes, encoding carboxylate transporters, which are important in the early stages of infection. This work investigated how host microenvironments, in particular acidic containing lactic acid, affect C. albicans biofilm formation and antifungal drug resistance. Multiple components of the extracellular matrix were also analysed, including their impact on antifungal drug resistance, and the involvement of both Jen1 and Jen2 in this process. The results show that growth on lactate affects biofilm formation, morphology and susceptibility to fluconazole and that both Jen1 and Jen2 might play a role in these processes. These results support the view that the adaptation of Candida cells to the carbon source present in the host niches affects their pathogenicity.This study was funded by the Portuguese Foundation for Science and Technology (FCT) [grant number PTDC/BIAMIC/5184/2014]. SM, CFR and RA received FCT PhD studentships [grant numbers SFRH/BD/74790/2010, SFRH/ BD/93078/2013, PD/BD/113813/2015, respectively]. The work on CBMA was supported by FCT [grant number UID/ BIA/04050/2013] and COMPETE 2020 [grant number POCI01-0145-FEDER-007569]. The work on CEB was supported by FCT [grant number UID/BIO/04469/2013], COMPETE 2020 [grant number POCI-01-0145-FEDER-006684] and BioTecNorte operation [grant number NORTE-01-0145FEDER-000004] funded by the ERDF under the scope of Norte2020 – Programa Operacional Regional do Norte. AJPB was funded by the UK Medical Research Council [grant number MR/M026663/1]; by the UK Biotechnology and Biological Research Council [grant number BB/K017365/1]; and by the MRC Centre for Medical Mycology and the University of Aberdeen [grant number MR/M026663/1]. The funders had no role in study design, data collection and analysis, the decision to publish, or in the preparation of the manuscript.info:eu-repo/semantics/publishedVersio

Laboratorial approach in the diagnosis of food allergy

OBJCTIVE: Review the available laboratory tests used to assist in the diagnosis of IgE-mediated and non-IgE-mediated food allergy. DATA SOURCES: Papers in English and Portuguese published in PubMed and Embase, in the last ten years. Terms searched were food allergy, diagnose and laboratory, isolated and/or associated. DATA SYNTHESIS: The diagnostic approach to food allergy reactions includes a good medical history, laboratory studies, elimination diets and blinded food challenges. More recently, the use of a quantitative measurement of food-specific IgE antibodies has been shown to be more predictive of symptomatic IgE-mediated food allergy. Food-specific IgE serum levels exceeding the diagnostic values indicate that the patient is greater than 95% likely to experience an allergic reaction if he/she ingests the specific food. Such decision point values have been defined just for some foods and inconsistent results were obtained when allergy to the same food was studied in different centers. Food challenges, in particular the double-blind placebo-controlled food challenge (DBPCFC), represent the most reliable way to establish or rule out food hypersensitivity. CONCLUSIONS: A number of recent developments are improving the predictive value of some laboratory tests for the diagnosis of food allergies. However, to date, no in-vitro or in-vivo test shows full correlation with clinical food allergy and the DBPCFC remains the gold standard for the definitive diagnosis of specific food allergies. There is an urgent need for new and fundamentally improved diagnostic approaches, which must be validated in patients with food allergy confirmed by a positive DBPCFC.OBJETIVO: Revisar os exames laboratoriais disponíveis utilizados no diagnóstico da alergia alimentar mediada ou não por IgE. FONTES DE DADOS: Artigos publicados em base de dados PubMed e Embase (língua inglesa e portuguesa) nos últimos dez anos. As palavras-chave utilizadas como fonte de busca foram alergia alimentar, diagnóstico e laboratório, isolados e/ou associados. SÍNTESE DOS DADOS: A abordagem diagnóstica das reações alérgicas a alimentos inclui história clínica completa, estudos laboratoriais, dietas de eliminação e desencadeamentos cegos com alimentos. Recentemente, a medida quantitativa de anticorpos IgE específicos a alimentos tem mostrado ser mais preditiva de alergia alimentar sintomática mediada por IgE. Níveis séricos de IgE específica a alimento que excedam os valores diagnósticos indicam que o paciente tem chance maior que 95% de apresentar uma reação alérgica se ingerir o alimento em questão. Estes valores de decisão foram definidos para alguns alimentos e resultados inconsistentes são obtidos ao se estudar diferentes populações. Os desencadeamentos com alimento, especialmente o duplo-cego controlado por placebo (DADCCP), representa a maneira mais confiável de estabelecer ou descartar o diagnóstico de hipersensibilidade alimentar. CONCLUSÕES: Número crescente de aquisições tem melhorado o valor preditivo de alguns testes laboratoriais empregados no diagnóstico de alergias alimentares. Entretanto, até hoje, não há teste in vitro ou in vivo que mostre correlação completa com a clínica da alergia alimentar. O DADCCP continua sendo o padrão-ouro no diagnóstico definitivo de alergia alimentar específica. São necessárias, urgentemente, novas abordagens diagnósticas válidadas em pacientes com alergia alimentar confirmada por DADCCP positivo.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de PediatriaUNIFESP-EPM Departamento de PediatriaUniversidade de São Paulo Faculdade de Medicina Departamento de PediatriaUniversidade Federal da Bahia Departamento de PediatriaUNIFESP-EPMUniversidade Federal do Paraná Departamento de PediatriaUNIFESP, EPM, Depto. de PediatriaUNIFESP, EPM Depto. de PediatriaUNIFESP, EPMSciEL

Ambient vibration tests of a cross-laminated timber building

Cross-laminated timber has, in the last 6 years, been used for the first time to form shear walls and cores in multi-storey buildings of seven storeys or more. Such buildings can have low mass in comparison to conventional structural forms. This low mass means that, as cross-laminated timber is used for taller buildings still, their dynamic movement under wind load is likely to be a key design parameter. An understanding of dynamic lateral stiffness and damping, which has so far been insufficiently researched, will be vital to the effective design for wind-induced vibration. In this study, an ambient vibration method is used to identify the dynamic properties of a seven-storey cross-laminated timber building in situ. The random decrement method is used, along with the Ibrahim time domain method, to extract the modal properties of the structure from the acceleration measured under ambient conditions. The results show that this output-only modal analysis method can be used to extract modal information from such a building, and that information is compared with a simple structural model. Measurements on two occasions during construction show the effect of non-structural elements on the modal properties of the structure

Identifying Implicit Vulnerabilities through Personas as Goal Models

When used in requirements processes and tools, personas have the potential to identify vulnerabilities resulting from misalignment between user expectations and system goals. Typically, however, this potential is unfulfilled as personas and system goals are captured with different mindsets, by different teams, and for different purposes. If personas are visualised as goal models, it may be easier for stakeholders to see implications of their goals being satisfied or denied, and designers to incorporate the creation and analysis of such models into the broader RE tool-chain. This paper outlines a tool-supported approach for finding implicit vulnerabilities from user and system goals by reframing personas as social goal models. We illustrate this approach with a case study where previously hidden vulnerabilities based on human behaviour were identified

Optimization and characterization of bacterial nanocellulose produced by Komagataeibacter rhaeticus K3

Supplementary material associated with this article can be found, in the online version, at doi:10.1016/j.carpta.2020.100022.In this work, a novel Bacterial NanoCellulose (BNC) producing strain, from Kombucha tea, was isolated and characterized. Based on 16S rRNA analysis the strain was identified as Komagataeibacter rhaeticus. Under static culture, K. rhaeticus K3 produces membranes with a relaxed structure, as observed by Scanning Electron Microscopy (SEM). The addition of 2% (v/v) ethanol to the culture media enhanced by more than 3-fold of the BNC yield. Response surface methodology (RSM) was performed with K. rhaeticus K3, using a new low cost Eucalyptus Biomass Hydrolysate (EBH). The maximum experimental BNC yield was of 5.46 g/L, obtained with the following composition: 31.4 g/L of EBH; 2.89% (v/v) of ethanol and 10.8 g/L of Yeast extract/peptone. Texture Profile Analysis (TPA) of BNC membranes obtained using Hestrin-Schramm culture (HS) medium and optimized medium from EBH showed that membranes from EBH had higher resistance to compression, higher cohesiveness and resilience.This study was supported with the funds of Institute of Technical Biochemistry, Lodz University of Technology, Lodz, Poland and from The Navigator Company through the I&D no. 21874, “Inpactus-– Produtos e Tecnologias Inovadores a partir do Eucalipto”, funded through the Fundo Europeu de Desenvolvimento Regional (FEDER) and the Programa Operacional Competitividade e Internacionalização (POCI) is greatly acknowledged. This study was also supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI01-0145-FEDER-006684) 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 financial support of the FCT (ESF) through the grant given to Francisco A.G. Soares da Silva (SFRH/BD/146375/2019).info:eu-repo/semantics/publishedVersio

Determinants on an efficient cellulase recycling process for the production of bioethanol from recycled paper sludge under high solid loadings

Background: In spite of the continuous efforts and investments in the last decades, lignocellulosic ethanol is still not economically competitive with fossil fuels. Optimization is still required in different parts of the process. Namely, the cost effective usage of enzymes has been pursued by different strategies, one of them being recycling. Results: Cellulase recycling was analyzed on Recycled Paper Sludge (RPS) conversion into bioethanol under intensified conditions. Different cocktails were studied regarding thermostability, hydrolysis efficiency, distribution in the multiphasic system and recovery from solid. Celluclast showed inferior stability at higher temperatures (45-55 ºC), nevertheless its performance at moderate temperatures (40ºC) was slightly superior to other cocktails (ACCELLERASE®1500 and Cellic®CTec2). Celluclast distribution in the solid-liquid medium was also more favorable, enabling to recover 88 % of final activity at the end of the process. A Central Composite Design studied the influence of solids concentration and enzyme dosage on RPS conversion by Celluclast. Solids concentration showed a significant positive effect on glucose production, no major limitations being found from utilizing high amounts of solids under the studied conditions. Increasing enzyme loading from 20 to 30 FPU/ gcellulose had no significant effect on sugars production, suggesting that 22 % solids and 20 FPU/gcellulose are the best operational conditions towards an intensified process. Applying these, a system of multiple rounds of hydrolysis with enzyme recycling was implemented, allowing to maintain steady levels of enzyme activity with only 50 % of enzyme on each recycling stage. Additionally, interesting levels of solid conversion (70-81 %) were also achieved, leading to considerable improvements on glucose and ethanol production comparatively with the reports available so far (3.4 and 3.8 fold, respectively). Conclusions: Enzyme recycling viability depends on enzyme distribution between the solid and liquid phases at the end of hydrolysis, as well as enzymes thermostability. Both are critical features to be observed for a judicious choice of enzyme cocktail. This work demonstrates that enzyme recycling in intensified biomass degradation can be achieved through simple means. The process is possibly much more effective at larger scale, hence novel enzyme formulations favoring this possibility should be developed for industrial usage.This work had the fnancial support of the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/ BIO/04469/2013 unit, COMPETE 2020 (POCI-01-0145-FEDER-006684) and the MultiBiorefnery project (POCI-01-0145-FEDER-016403). Furthermore, FCT equally supported the Ph.D. grant to DG (SFRH/BD/88623/2012).info:eu-repo/semantics/publishedVersio