Translational Research and Innovation in Human and Health Science

AFM as a tool to evaluate the risk for cardiovascular diseases in patientsCONTACT Nuno Santos [email protected]

Abstract

Erythrocyte aggregation is an indicator of cardiovascular risk, which is influenced by plasma fibrinogen concentration. Fibrinogen levels are elevated during cardiovascular diseases. Our main goals were to understand how fibrinogen-erythrocyte binding influences erythrocyte aggregation and how it constitutes a cardiovascular risk factor in essential arterial hypertension (EAH) and chronic heart failure (CHF). Fibrinogen-erythrocyte and erythrocyte-erythrocyte adhesion measurements were conducted by atomic force microscopy (AFM)-based force spectroscopy. Upon increasing fibrinogen concentration, there was an increase in the work and force necessary for cell-cell detachment, both for healthy donors and EAH patients. Nevertheless, higher values were obtained for the EAH patients at each fibrinogen concentration. Fibrinogen-erythrocyte (un)binding forces were higher in EAH and CHF patients, when compared with the control group, despite a lower binding frequency. Ischemic CHF patients showed increased binding forces compared to non-ischemic patients. Erythrocyte deformability (assessed as elongation index) results show that heart failure patients presented higher erythrocyte deformability than the control group at lower shear stresses, and lower deformability at higher shear stresses. This indicates that patients’ erythrocytes are more deformable than those from healthy donors in blood vessels with larger internal diameters; however, in smaller-diameter vessels the opposite trend exists. Finally, a 12-month clinical follow-up shows that CHF patients with higher fibrinogen-erythrocyte binding forces, probed by AFM at the beginning of the assessment, had a significantly higher probability of being hospitalized due to cardiovascular complications on the subsequent year. Our results show that AFM can be a promising tool for clinical prognosis, pinpointing those patients with increased risk for cardiovascular diseases.

CONTACT Nuno Santos [email protected]

Gla-rich protein, a vitamin K-dependent protein involved in inflammation and calcification-related [email protected]

Abstract

Inflammation and calcification-related diseases are pathological processes involving several factors and a complex interplay between inflammation and calcification events that lead to disease progression. Gla-rich protein (GRP) is a vitamin K dependent protein (VKDP) (1) shown to function as a calcification inhibitor in cardiovascular [2] and articular tissues [3] and as an anti-inflammatory agent in chondrocytes, synoviocytes and monocytes/macrophages [2,3,4]. In a context of chronic inflammation and calcification-related pathologies, GRP is able to act as an endogenous mediator of inflammatory responses and a novel molecular mediator linking inflammation and calcification events, with potential therapeutic application (4). Vascular calcification (VC) is highly prevalent in chronic kidney disease (CKD) and associated with increased morbidity and mortality. This complex process, known to be associated with inflammatory status and enhanced by inflammatory cytokines, relies on the presence of systemic and local calcification inhibitors like fetuin-A and Gla-rich protein (GRP). Increased levels of calcium (Ca), and Ca phosphate product in circulation have been related with increased frequency of VC in CKD. The study of circulating nanoparticles as calciprotein particles (CPPs) and extracellular vesicles (EVs) and their pathophysiological role and mechanistic link, with the prevalence of vascular calcification in CKD patients, is greatly highlighted. In particular, results on the association of GRP with CPP and circulating EVs isolated from CKD patients show a higher mineral content and reduced levels of GRP and fetuin A, when compared with healthy controls. Moreover, the study of the effect of these biological CKD nanoparticles in VSMC calcification, differentiation and inflammation, reveal its pro-inflammatory and mineralization pathogenic activity. In conclusion, in a context of chronic inflammation and calcification-related pathologies, GRP acts as a novel molecular mediator linking inflammation and calcification events, with potential therapeutic application. This also precludes a great potential use for GRP, as part of circulating EVs and CPPS, for the diagnostic of these highly prevalent vascular calcification-related diseases.

Acknowledgements: This work was funded by projects PTDC/SAU-ORG/117266/2010, PTDC/BIM-MEC/1168/2012 and project UID/Multi/04326/2013, all from the Portuguese Science and Technology Foundation (FCT) and UCIBIO-REQUIMTE.

CONTACT Dina Simes [email protected]

Aquaporins are novel targets for obesity and [email protected]

Abstract

Aquaporins (AQPs) are membrane channels that transport water and glycerol across cell membranes and are crucial for many biological functions. Mammals AQPs (0-12) have essential roles in health and disease, suggesting that their modulation may have broad therapeutic potential. Detecting AQPs expression and function in cells and tissues is critical for screening modulators for the development of efficient medicines [1]. We have disclosed inhibition of the aquaglyceroporin AQP3 by gold-based compounds [2] and highlighted the relation between AQP3 expression, glycerol permeation across cell membranes and cell proliferation [3]. The mechanism of inhibition has been recently described using molecular dynamics, combined with density functional theory and electrochemical studies [4].

The selectivity of gold compounds towards aquaglyceroporins was confirmed by its inhibitory effect on AQP7, largely expressed in adipocytes and important in adipose tissue homeostasis and obesity [5]. By facilitating glycerol efflux from adipocytes during fasting and its uptake into the liver for gluconeogenesis, aquaglyceroporins are emerging as key players in adipose tissue homeostasis and insulin response with potential implications in obesity and metabolic-related complications [6].

AQPs are also involved in tumor invasion, metastasis and growth [7]. Our recent studies pointed to a role of AQP5 on adipocyte cell differentiation [8] and its involvement in cell oxidative stress response by facilitating hydrogen peroxide permeation across membranes [9]. Interestingly, we have also detected aberrant expression of AQP5 in pancreatic tumors of high malignancy, suggesting it may be used as a biomarker for early diagnosis [10].

Altogether, our studies highlight AQPs as promising drug targets in obesity and cancer.

CONTACT Graça Soveral [email protected]

Paradigm of antibiotic resistant: old or new?

Abstract

Nosocomial infections caused by multidrug-resistant bacteria represent a growing problem worldwide and are well documented. There are statements by the WHO and CDC describing a global crisis and an impending catastrophe of a return to the pre-antibiotic era. Metagenomic studies have revealed homologues of known resistance genes broadly distributed across environmental locales and allowed the formation of a database with more than 20,000 potential resistance genes predicted from available bacterial genome sequences. However, the number of resistance determinants functional in pathogens are much smaller. The widespread dissemination of antibiotic resistance elements is inconsistent with a hypothesis of contemporary emergence and instead suggests a richer natural history of resistance. Several mechanisms are responsible for the emergence and prevalence of antimicrobial resistance, and such mechanisms have been divided into genetic mutation occurred at a low frequency and acquisition of various genes among bacteria. All the known antibiotic resistance mechanisms, acquired by opportunistic and pathogenic bacteria, evolve mutations occurring in pre-existing genes of the bacterial chromosome positively selected by environmental forces and can be responsible for the decreased affinity of antibiotics to their targets. Acquisition of resistance genes has been regarded as major contributor for the wide distribution and spread of antimicrobial resistance, via either vertical transfer or horizontal transfer, with the latter mechanism involving mobile genetic elements such as plasmids, transposons, integrons and bacteriophages. The role played by these elements among bacteria has been well established and documented, which had also been considered to contribute to the unleashing of “Super Bugs”. However little is known about the diversity and origin of resistance genes among the unculturable environmental bacteria. The soil is one potentially rich but largely unstudied reservoir. The complexity of its microbial community coupled with its high density of antibiotic-producing bacteria makes the soil a likely origin for diverse antibiotic resistance determinants. This raises many questions about the natural functions of environment. Recently, there are evidences that environmental habitats especially waterbodies such as rivers and streams are ideal vectors for the antibiotic resistance dissemination. The emergence and rise of antibiotic resistance observed worldwide cannot be explained only by the increasing modern use of antibiotics in humans, but involves a complex interaction in an ecosystem comprising microbial communities, antibiotics, and antibiotic resistance genes.

CONTACT Aida Duarte [email protected]

The impact of the presence of bacteria on dental materials [email protected]

Abstract

The human oral cavity holds one of the most diverse microbiome in the human body. It includes viruses, fungi, protozoa, archaea and bacteria. These microorganisms regulate each other and coexist within the microbial community. They can be found in complex biofilms attached to soft and hard tissues assembled in an ordered process that begins with adherence of microbial colonizers to a surface followed by attachment of different microbial species to initial colonizers. The presence of these microorganisms may affect the wear.

In order to understand the impact of the presence of bacteria on the dental materials testing two tribological systems were studied: zirconia dental crown against natural teeth and zirconia abutment against titanium implant. Thus in order to evaluate the effect of the presence of bacteria a nom cariogenic bacteria was selected: Streptococcus salivarius (S. salivarius). The bacteria growth profiles were studied in artificial saliva with three different nutrients (meat extract, urea and glucose), pH (from 5 to 9) and motion conditions (static and movement) in order to set the wear tests.

Reciprocating pin on plate tests were performed for both tribological systems (human cusps/zirconia plates and Ti6Al4V ball/zirconia plates) using three lubricants: artificial saliva, artificial saliva with glucose and artificial saliva with glucose and S. salivarius, at a temperature of 37 °C and pH 7.

The best bacteria growth condition were found for glucose as nutrient, at pH 7 and without motion.

For the tribological system human cusps/zirconia plates the results showed that the presence of bacteria reduced the friction coefficient, dental wear loss and reduces de thickness of the smear layer formed on the worn surfaces.

For the tribological system Ti6Al4V ball/zirconia plates the results revealed that bacteria reduced the titanium wear and the titanium corrosion activity during wear.

Over all, the presence of S. salivarius leads to a dramatic changes of the tribological performance of the tested dental materials.

Acknowledgements: The authors are grateful to Fundação para a Ciência e Tecnologia (FCT, Portugal) for financial support under project Pest-OE/CTM/UI0084/2014.

CONTACT Célio Figueiredo-Pina [email protected]

Zinc is a bioactive element in restorative DentistryCONTACT [email protected]

Abstract

To promote resin polymers adhesion to dentin, the mineral phase from the substrate has to be removed by acids and the voids left by mineral should be filled with the adhesive resin that undergoes complete in situ polymerization to form the hybrid layer. It is generally accepted that there is a discrepancy between the depth of demineralization and the depth of resin infiltration. Matrix metalloproteinases are present in dentin and contribute to collagen degradation at the dentin–resin bonded interfaces, jeopardizing the longevity of bonded restorations. A decrease of bonding efficacy over time has been reported and attributed to the action of host-derived MMPs. Therefore, one of the proposed strategies used to increase the longevity of adhesive restorations has been to inhibit the MMP-induced degradation of dentin collagen [1,2]. Zinc effectively reduces the MMP-mediated degradation of collagen in dentin [1,2]. It seems that subtle conformational changes occur in collagen following zinc binding and lead to the protection of sensitive cleavage sites of metalloproteinases. Moreover, zinc is able to promote dentin remineralization [3,4]. Zinc and phosphate are important for hydroxylapatite homeostasis. Scholzite formation was encountered in dentin stored in zinc-containing solutions [3]. Zinc might allow to reach the balance between dentin demineralization and remineralization processes. Thus, the zinc concentration around matrix-bound MMPs might be very important in determining longevity of resin-dentin interfaces [4].

Acknowledgements: This work was supported by the Ministry of Economy and Competitiveness (MINECO) [Project MAT2014-52036-P] and European Regional Development Fund (FEDER).

CONTACT Manuel Toledano-Osorio [email protected]

Polymers as nanocarriers for dental therapyCONTACT [email protected]

Abstract

Introduction: The aim of this study was to evaluate the effectiveness of different nanogels-based solutions for dentin permeability reduction and to determine the viscoelastic performance of cervical dentin after their application.

Materials and Methods: Four experimental nanogels based on zinc, calcium or doxycycline-loaded polymeric nanoparticles (four products) were applied on citric acid etched dentin (fifteen specimens) to facilitate the occlusion and the reduction of the fluid flow at the dentinal tubules. After 24 h and 7 d of storage, cervical dentin was evaluated by a highly sensitive fluid filtration system. Field emission scanning electron microscopy, energy dispersive analysis, AFM and Nano-DMA analysis were performed. Complex, storage, loss modulus and tan delta (δ) were assessed [1].

Results: Tubules were 100% covered in dentin treated with Ca-nanoparticles and Zn-nanoparticles, analysed at 24 h and 7 d, respectively. All tubules were visible after 7 d when dentin was treated with Ca-NPs, and 95% of them were filled. Dentin treated with both Zn-nanoparticles and Ca-nanoparticles attained the greatest reduction of dentinal fluid flow. The complex modulus and the tan delta between peritubular and intertubular dentin treated with Zn-nanoparticles was bigger than that obtained with when Ca-nanoparticles, showing zones of stress concentrations with dentin structure breakdown [2].

Discussion and Conclusions: Crack-bridging with frictional pullout due to precipitation of minerals were produced. This mineralization, as sight of energy dissipation, improved the sealing of the micro-cracks and the hydraulic conductance of dentin [2,3].

Acknowledgements: This work was supported by the Ministry of Economy and Competitiveness (MINECO) [Project MAT2014-52036-P] and European Regional Development Fund (FEDER).

CONTACT Manuel Toledano-Osorio [email protected]

Can masticatory function contribute to a fast facial temperature recovery? A comparative study based on thermography evaluatione-mail of the corresponding author: [email protected]

Abstract

Introduction: Thermography is a non-radiating and contact-free technology which can be used to monitor skin temperature. The efficiency and safety of thermography make it a useful tool for detecting and locating thermal changes in skin surface. Being a non-invasive technology, the interest of its application in the field of health has been growing, with its application in biomechanics domains. This work intends to be a contribution for the use of thermography as a methodology for evaluation of skin temperature in the context of orofacial biomechanics. The study aims to identify the oscillations of skin temperature in the left and right hemiface regions of the masseter muscle, before, during and after ice therapy. The study involves the observation of the recovery with and without an induced stimulus based on the use of a chewing gum.

Materials and Methods: Using a FLIR T430sc camera, a data acquisition protocol was followed with a group of 8 volunteers, aged between 22 and 27 years. The tests were performed in a controlled environment with the volunteers in a comfortably static position. The thermal stimulus involves the use of an ice volume with controlled size and contact surface. The skin surface temperature was recorded in two distinct situations, namely without further stimulus and with the addition of the induced stimulus. The thermal data obtained were treated using FLIR Research IR Max software. The considered emissivity was set to 0.98, the normal value for the skin [3]. The statistical analysis of the results was performed with RStudio Software.

Results: The results obtained show that the time required to recover until the initial temperature ranged from 20 to 52 min when no stimulus was added and ranged between 8 and 26 min with the chewing gum stimulus. The analysis of the results reveals a variation of averages with statistical significance when comparing the application and absence of stimulus with a p = .4756 (p > .05).

Discussion and conclusions: This work, despite the reduced sample, intends to demonstrate the utility of thermography in the study of some aspects of orofacial biomechanics. In fact, the applied methodology clearly shows a decrease in the time needed to reestablish the initial temperature when implementing a simple chewing gum stimulus. This can be due to the increase in metabolic recruitment of the muscle by the chewing process, with an increase of blood circulation on muscle metabolism and by the neutralizing effect of the vascularization itself increased by the function. In other hand, dentists should be aware that with the current ice protocols they are most likely to be maintaining skin temperature in lower values for longer times, raising questions of potential iatrogenic effects and tissue hypothermia by maintaining a temperature much lower than normal values. However, these questions remain unclear and not studied in the literature.

Acknowledgements: The authors would like to thank for LBA – Applied Biomechanics Laboratory from Polytechnic Institute of Coimbra and also all volunteers of the study. This research was sponsored by the project UID/EMS/00285/2013.

CONTACT Ana Dionísio [email protected]

25I-NBOH: analytical identification of a new potent serotonin 5-HT2A receptor agonist

Abstract

Introduction: The last decade presented an unprecedented increase in the number of new psychoactive substances (NPS) released in the drug market [1]. Recently a new class of drugs known as NBOH started to be reported in many Brazilian States [2]. The NBOH compounds are N-benzylhydroxy derivatives of the 2C hallucinogen. The 25I-NBOH is a liable molecule that undergoes degradation when exposed to routine screening gas chromatography methods posing an extra challenge to correct identification as the main degradation product is another phenethylamine, 2C-I [2].

Materials and Methods: GC–MS analyses were performed on an Agilent 7890A gas chromatograph connected to a 5975C Mass Spectrometer (Agilent Technologies, Santa Clara, CA, USA.). An Agilent J&W HP-1MS fused silica capillary column was used. Sample injection volume was 1 μL with a 25:1 split ratio. Helium was used as carrier gas and injector temperature was 280 °C. The oven program: t 150 °C for 1.5 min, 30 °C min⁻¹ to 250 °C hold for 1 min, and 50 °C min⁻¹ to 300 °C hold for 3 min. Mass scan range was m/z 35–550.

Results: Firstly, we present the GC–mass spectrometry (MS), liquid chromatography–quadrupole time-of-flight-MS, and Fourier transform infrared and nuclear magnetic resonance analyses to complete 25I-NBOH molecular characterization. Secondly, a GC-MS method that allows distinction between 25I-NBOH and 2C-I using routine GC–MS without resorting to derivatization is described. While direct detection of 25I-NBOH under routine GC conditions is still to be achieved, slight adjustments in standard GC methods, including shortening of the solvent delay window, enabled the detection of an additional peak containing 25I-NBOH degradation product’s fragmentation ions. Consequently, this secondary early chromatographic peak is characteristic of the 25I-NBOH, thus preventing misidentification.

Discussion and Conclusions: With the active and continuous emergence of NPSs over the last years, most of them derived from previously known ones, correct identification of questioned samples becomes an increasingly subtler task, because some species of compounds undergo chemical changes during analysis, potentially leading to serious misidentification. In this context, forensic scientists dealing with routine GC–MS testing of seized samples should be mindful of this issue and be ready to recognize the possibility that such a change may take place during analysis, specially when solely relying on GC–MS.³

CONTACT Ana Flávia Belchior de Andrade [email protected]

Occupational stress among Portuguese journalists: a comparative study between reporting daily and critical eventse-mail of the corresponding author: [email protected]

Abstract

Introduction: Nowadays, journalism is one of the top ten most stressful occupations [1] but it was only after 9/11, that research has come to acknowledge journalists as an at-risk occupational group, and to focus on the impact of the events they report on their health and well-being. A recent meta-analysis of studies from 2001 to 2015 on journalists’ occupational stress revealed that little is known about the differences and similarities between occupational stress experiences and variables of journalists in their daily work and in the context of reporting critical events [2]. Furthermore, there are no studies that try to identify the positive impacts on the life and health of these professionals and that use a theoretical occupational model as a backdrop to their research. Taking the occupational stress model [3] as a framework, this study attempts to overcome this shortcomings by seeking to shed light upon the similarities and differences in occupational stress variables between daily work journalists and special envoys reporting major disasters, thus offering important contributions to theory and practice. This study also brings together both qualitative and quantitative data in the analysis of the variables under study, according to the number of deployments to a critical event.

Method: 25 Portuguese journalists were interviewed on their perceptions of the core variables of the model [3]: occupational stressors, emotional reactions, coping and savoring strategies and the consequences of these experiences on their well-being, either in their daily context or critical scenarios. The number of times they had been abroad reporting a major disaster (natural or human made) was also questioned in order to ascertain whether repeated exposure to trauma influenced journalists’ occupational stress perceptions.

Results: The data content analysis gave rise to 113 inter-related categories, structured in a hierarchical system. For the daily professional context of the journalists, 39 categories (35%) were identified while for critical events, 74 categories (65%) were constructed. Occupational stressors and emotional reactions differed across both settings, emotion-focused coping strategies were the most frequently mentioned for both settings, while savoring strategies were only referred to for critical events. Perceptions of coping strategies were four times more frequent than those of savoring. Consequences associated with journalists’ experiences were perceived as being mainly negative in both occupational contexts. Significant differences were identified in all of these variables according to the number of deployments to a critical event.

Discussion and conclusions: This study offers a valid contribution to the understanding of occupational stress among journalists and it serves to alert media organizations to the fact that suitable preparation and support should be provided to journalists in their work performance.

Funding: This work was supported by the Fundação para a Ciência e Tecnologia of Portugal [grant number SFRH/BD/41454/2007].

CONTACT Susana Monteiro [email protected]

The return to everyday life after experiencing a limit situation

Abstract

In this study we try to understand the experience of the return to everyday life of adults faced with a limit-situation; to make clear the experienced processes involved in the limit-situation and to describe the lived circumscribed experience and punctuating the return path to everyday life.

It is based in the health existential concept within the framework of caring as a vital dimension of human behaviour, namely when the challenge is to safeguard and activate all that makes living and existence possible. The experience of the concrete possibility and close to finitude is a process present in the human response facing the disruption of a serious illness lived as a limit-situation.

The phenomenological approach has permitted access to the narration of the lived experience of returning to everyday life. The meaning units are determined by the experienced moments – the way the subject relates to them, lives them in its own body and inscribes in the grammar of his/her existence – that trace them as analytical possibilities in the context of this study.

The event is a threatening situation for the person due to the suffering, unease and strangeness experienced suddenly and expressed in the body, now the focus of all attention. The body is an object outside oneself, revealing the division body-consciousness. The event it triggers is not lived as an illness but as a struggle between death and life, each participant searching for meaning and reason in the context of existence. It is a threat to concrete survival, to the physical body, biologic in its vital functions. In this perspective, the return to everyday life, its existence in (his/her) world is seen as a “hard” work of surviving the fight it has begun, and, as it is solitary work, it lacks the presence and companionship of others. It is a transformation work of the self – to become another, better person. At first it is the work of delivering oneself from the disease in the body which is followed by silent and long work of freeing the disease from life which in turn translates into trusting the body that you are and “drop the illness’s routines, enter the everyday life circuit, and this is what best characterizes life and feeling healthy within it”.

Keywords:

• limit-situation
• lived experience
• nursing
• return to everyday life

CONTACT Maria Teresa Rebelo [email protected]

Organizational environment of nursing professional practiceE-mail contact: [email protected]

Abstract

Introduction: Nursing practice care has been developed in an environment of increasing complexity [1]. Research shown that environment health organizations should be favorable to nursing professional practice and is fundamental to improve quality care provided. Many studies identify some traits of the organizational environment in which care are provided and influence client’s outcomes, nurses, and health organizations. Positive nursing practice environments are associated with better perceptions of nurses' quality of care and satisfaction [2,3]. The aim of this study was to evaluate nursing environment professional practice in four public portuguese hospitals.

Materials and Methods: This study is a quantitative, observational, descriptive, cross-sectional study, with the research question: How nurses evaluate the organizational environment professional practice? To evaluate the nurses professional environment was applied the Portuguese version of the Revised Nursing Work Index (NWI-R). Ethical procedures such as informed consent have been followed. Of the 1304 nurses of four public portuguese hospitals, selected for the study who were invited, 767 nurses collaborated, responding to the questionnaires.

Results: The nurses who participated in the study 83.4% are licensed, 84.2% female, with an average age of 37.24 years and 14.25 years of professional activity, 47.4% were working in medical units, 7.7% in Surgery and 44.9% in units of other types. The nursing professional practice environment had a generally positive assessment (mean of 3.33). All subscales that are defined in the study of Aiken [4], Multidisciplinary Relation, Autonomy, Organizational Support and Control over the Environment - had positive averages with 3.55; 3.35; 3.30 and 3.12, respectively.

Discussion and Conclusions: A professional nursing organizational environment was generally considered favorable. The Control over the Environment, Organizational Support and Autonomy are strongly conditioning of a conductive environment to the practice of nursing care. Contributions to consider to Nursing Management and Nursing Research should be considered.

CONTACT Teresa Ramalhal [email protected]