Combined Application of Antisense Oligomers to Control Transcription Factors of Candida albicans Biofilm Formation.

Antisense oligomers (ASOs) have been little exploited to control determinants of Candida albicans virulence. Biofilm formation is an important virulence factor of C. albicans, that is regulated by a complex network of transcription factors (such as EFG1, BRG1 and ROB1). Thus, the main goal of this work was to project ASOs, based on the 2'-OMethyl chemical modification, to target BRG1 and ROB1 mRNA and to validate its application either alone or in combination with the EFG1 mRNA target, to reduce C. albicans biofilm formation. The ability of ASOs to control gene expression was evaluate by qRT-PCR. The effect on biofilm formation was determined by the total biomass quantification, and simultaneously the carbohydrates and proteins reduction on extracellular matrix. It was verified that all the oligomers were able to reduce the levels of gene expression and the ability of C. albicans to form biofilms. Furthermore, the combined application of the cocktail of ASOs enhances the inhibition of C. albicans biofilm formation, minimizing biofilm thickness by reducing the quantity of matrix content (protein and carbohydrate). So, our work confirms that ASOs are useful tools for research and therapeutic development on the control of Candida species biofilm formation.

Deep brain stimulation of the dorsal raphe induces anxiolytic and panicolytic-like effects and alters serotonin immunoreactivity.

Previously we showed that Deep Brain Stimulation (DBS) of the dorsal region (DRD) and of the lateral wings of the dorsal raphe (lwDR) respectively decreases anxiety and panic-like responses in the elevated T-maze (ETM). This study investigates neurobiological alterations which might respond for these behavioral effects. Male Wistar rats were submitted to high-frequency stimulation (100 µA, 100 Hz) of the DRD or of the lwDR for 1 h, and subsequently tested in the avoidance or escape tasks of the ETM. Since serotonin (5-HT) reuptake inhibitors are first line pharmacological treatment for anxiety disorders, we also tested the effects of chronic fluoxetine administration (10 mg/kg, IP, 21 days) on a separate group of rats. An open field was used for locomotor activity assessment. Additionally, we evaluated c-Fos immunoreactivity (Fos-ir) in serotonergic cells of the dorsal raphe (DR). Results showed that DBS of the DRD decreases avoidance reactions, an anxiolytic-like effect, without altering escape or locomotor activity. Both fluoxetine and DBS of the lwDR decreased escape responses in the ETM, a panicolytic-like effect, without altering avoidance measurements or locomotor activity. While DBS of the DRD decreased double immunostaining in the DRD, DBS of the lwDR increased Fos-ir and double immunostaining in the DRD and lwDR. Fluoxetine also increased double immunostaining in the lwDR and in the DRV but decreased it in the DRD. These results suggest that both the anxiolytic and panicolytic-like effects of DBS and fluoxetine are related to 5-HT modulation in different subnuclei of the DR.

ITER collective Thomson scattering-Preparing to diagnose fusion-born alpha particles (invited).

The ITER Collective Thomson scattering (CTS) diagnostic will measure the dynamics of fusion-born alpha particles in the burning ITER plasma by scattering a 1 MW 60 GHz gyrotron beam off fast-ion induced fluctuations in the plasma. The diagnostic will have seven measurement volumes across the ITER cross section and will resolve the alpha particle energies in the range from 300 keV to 3.5 MeV; importantly, the CTS diagnostic is the only diagnostic capable of measuring confined alpha particles for energies below ∼1.7 MeV and will also be sensitive to the other fast-ion populations. The temporal resolution is 100 ms, allowing the capture of dynamics on that timescale, and the typical spatial resolution is 10-50 cm. The development and design of the in-vessel and primary parts of the CTS diagnostic has been completed. This marks the beginning of a new phase of preparation to maximize the scientific benefit of the diagnostic, e.g., by investigating the capability to contribute to the determination of the fuel-ion ratio and the bulk ion temperature as well as integrating data analysis with other fast-ion and bulk-ion diagnostics.

Metal-based antimicrobial strategies against intramacrophage Mycobacterium tuberculosis.

The combinatorial chemistry has been an important tool for the development of new strategies against the Mycobacterium tuberculosis. Therefore, we evaluated the antimycobacterial activity of two coordinated metal complexes (Cu(II) and Co(II)) and a free ligand, including in the intramacrophage environment. The complexes were more active than the free ligand, indicating that the complexation favoured the antimicrobial activity. None of the compounds showed cytotoxic effect at the concentration of 200 µg ml-1 and both complexes showed intracellular antimicrobial activity, with results as effective as rifampicin. In this study, it was possible to identify complexes containing benzohydroxamate associated with transition metal ions (Cu2+ and Co2+ ), which were able to inhibit the growth of M. tuberculosis, including in persistence stage. In addition, the docking analysis allows inferring a possible interaction of the metal complexes with the enzyme urease, which has been reported as crucial for the bacillus survival in the intraphagosomal environment. Thus, these set of results demonstrate the potential of these metals in the development of new drugs against M. tuberculosis. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, it was possible to identify complexes containing benzohydroxamate associated with transition metals (Cu2+ and Co2+ ), which were able to inhibit the growth of Mycobacterium tuberculosis, including in the persistence stage. In this context, cobalt and copper can be scaffolds for new drugs against M. tuberculosis.

Metformin-associated lactic acidosis: A case reporting a serious complication in the perioperative period. / Acidosis láctica asociada a metformina: un caso clínico que reporta una complicación grave en el período perioperatorio.

The worldwide prevalence of diabetes mellitus (DM) has risen dramatically over the past two decades and this affects 10-15% of the surgical population1. Metformin represents the mainstay of anti-hyperglycaemic therapy for type 2 DM, providing numerous beneficial effects. However, a rare but severe adverse effect is the development of metformin-associated lactic acidosis (MALA) with high mortality rate. We describe a severe case of MALA in the postoperative period that culminated in cardiac arrest. Whereby anaesthetists should be knowledgeable and skilled in the care of patients with diabetes being imperative to focus on preoperative assessment, optimisation, management of pre-existing treatment during the starvation period and identification of patients medicated with metformin at risk of developing MALA.

Portuguese consensus document for the management of alpha-1-antitrypsin deficiency.

Alpha-1-antitrypsin deficiency (AATD) is a genetic autosomal codominant disorder caused by mutations in SERPINA1 gene. It is one of the most prevalent genetic disorders, although it remains underdiagnosed. Whereas at international level there are several areas of consensus on this disorder, in Portugal, inter-hospital heterogeneity in clinical practice and resources available have been adding difficulties in reaching a diagnosis and in making therapeutic decisions in this group of patients. This raised a need to draft a document expressing a national consensus for AATD. To this end, a group of experts in this field was created within the Portuguese Pulmonology Society - Study group on AATD, in order to elaborate the current manuscript. The authors reviewed the existing literature and provide here general guidance and extensive recommendations for the diagnosis and management of AATD that can be adopted by Portuguese clinicians from different areas of Medicine. This article is part of a supplement entitled "Portuguese consensus document for the management of alpha-1-antitrypsin deficiency" which is sponsored by Sociedade Portuguesa de Pneumologia.

Low concentrations of glyphosate-based herbicide cause complete loss of sperm motility of yellowtail tetra fish Astyanax lacustris.

Environmental relevant concentrations of glyphosate-based herbicide as 50 µg l-1 , 300 µg l-1 and 1800 µg l-1 can affect sperm quality of yellowtail tetra fish Astyanax lacustris. Viability of sperm cells was impaired at 300 µg l-1 , a concentration that is within legal limits in U.S.A. waterbodies, while motility was impaired at 50 µg l-1 , which is the more stringent limit set in Brazilian law. Therefore, environment protection agencies must review regulations of glyphosate-based herbicides on water bodies.

Towards large-scale in free-standing graphene and N-graphene sheets.

One of the greatest challenges in the commercialization of graphene and derivatives is production of high quality material in bulk quantities at low price and in a reproducible manner. The very limited control, or even lack of, over the synthesis process is one of the main problems of conventional approaches. Herein, we present a microwave plasma-enabled scalable route for continuous, large-scale fabrication of free-standing graphene and nitrogen doped graphene sheets. The method's crucial advantage relies on harnessing unique plasma mechanisms to control the material and energy fluxes of the main building units at the atomic scale. By tailoring the high energy density plasma environment and complementarily applying in situ IR and soft UV radiation, a controllable selective synthesis of high quality graphene sheets at 2 mg/min yield with prescribed structural qualities was achieved. Raman spectroscopy, scanning electron microscopy, high resolution transmission electron microscopy, X-ray photoelectron spectroscopy and Near Edge X-ray-absorption fine-structure spectroscopy were used to probe the morphological, chemical and microstructural features of the produced material. The method described here is scalable and show a potential for controllable, large-scale fabrication of other graphene derivatives and promotes microwave plasmas as a competitive, green, and cost-effective alternative to presently used chemical methods.

Effects of different sources of Saccharomyces cerevisiae biomass on milk production, composition, and aflatoxin M1 excretion in milk from dairy cows fed aflatoxin B1.

The aim of the present study was to evaluate the effect of different sources of Saccharomyces cerevisiae (SC) biomass (20.0 g/d) obtained from sugarcane (cell wall, CW; dried yeast, DY; autolyzed yeast, AY) and the beer industry (partially dehydrated brewery yeast, BY) on milk production, fat and protein percentages, and aflatoxin M1 (AFM1) excretion in milk from dairy cows receiving 480 µg aflatoxin B1 (AFB1) per day. A completely randomized design was used with 2 lactating cows assigned to each of 10 dietary treatments, as follows: negative controls (no AFB1 or SC-based biomass), positive controls (AFB1 alone), DY alone, DY + AFB1, BY alone, BY + AFB1, CW alone, CW + AFB1, AY alone, and AY + AFB1. The cows in the aflatoxin treatment group received AFB1 from d 1 to 6, while the SC biomass was administered with the AFB1 bolus from d 4 to 6. Aflatoxin B1 or SC-based products did not affect milk production or milk composition during the experimental period. Aflatoxin M1 was detected in the milk from all aflatoxin treatment group cows, reaching maximum levels at d 3 and varying from 0.52 ± 0.03 to 1.00 ± 0.04 µg/L. At end of the treatment period, CW, AY, DY, and BY removed 78%, 89%, 45%, and 50% of AFM1 from the milk, respectively, based on the highest level found on d 3. Results indicate a potential application of industrial fermentation by-products, especially CW and AY, as a feed additive in the diets of dairy cows to reduce the excretion of AFM1 in milk.

Performance of ultra-low-dose CT for the evaluation of coronary calcification: a direct comparison with coronary calcium score.

AIM: To evaluate the diagnostic performance of ultra-low-dose computed tomography (ULDCT) in comparison to standard coronary calcium score (CCS) acquisition for the evaluation of coronary artery calcification (CAC). MATERIALS AND METHODS: Standard CCS acquisition and ULDCT were performed in patients referred for coronary CT angiography for the evaluation of coronary artery disease. CAC in ULDCT was graded subjectively using a four-point scale (from 0, no calcification, to 3, severe calcification) for the complete study and for each individual coronary segment. The summation of all individual coronary segment scores generated an ULDCT total CAC score. ULDCT results were compared to standard Agatston score and sensitivity and specificity of ULDCT were calculated. RESULTS: CCS and ULDCT were performed in 74 patients, with a mean DLP of 77.7 mGy·cm (±12.1) and 9.3 mGy·cm (±0.6), respectively (p<0.001). Coronary calcification was detected in 47 patients (63.5%) in standard CCS acquisition (median Agatston score of 41; interquartile range [IQR]:0263), in comparison to 42 patients (56.8%) in ULDCT (p<0.001). The sensitivity and specificity of the ULDCT total CAC score ≥1 was 80.9% and 85.2%, respectively, with an accuracy of 82.4%. The area under the receiver operating characteristic curve for the presence of CAC was 0.87. CONCLUSION: ULDCT shows good sensitivity, specificity, and overall accuracy for the detection of coronary calcification with a markedly lower radiation dose in comparison to CCS. ULDCT is unlikely to miss coronary calcification in individuals with at least moderate calcium load (Agatston score >100).

Thermal conductivity as influenced by the temperature and apparent viscosity of dairy products.

This study aimed to evaluate the rheological behavior and thermal conductivity of dairy products, composed of the same chemical components but with different formulations, as a function of temperature. Subsequently, thermal conductivity was related to the apparent viscosity of yogurt, fermented dairy beverage, and fermented milk. Thermal conductivity measures and rheological tests were performed at 5, 10, 15, 20, and 25°C using linear probe heating and an oscillatory rheometer with concentric cylinder geometry, respectively. The results were compared with those calculated using the parallel, series, and Maxwell-Eucken models as a function of temperature, and the discrepancies in the results are discussed. Linear equations were fitted to evaluate the influence of temperature on the thermal conductivity of the dairy products. The rheological behavior, specifically apparent viscosity versus shear rate, was influenced by temperature. Herschel-Bulkley, power law, and Newton's law models were used to fit the experimental data. The Herschel-Bulkley model best described the adjustments for yogurt, the power law model did so for fermented dairy beverages, and Newton's law model did so for fermented milk and was then used to determine the rheological parameters. Fermented milk showed a Newtonian trend, whereas yogurt and fermented dairy beverage were shear thinning. Apparent viscosity was correlated with temperature by the Arrhenius equation. The formulation influenced the effective thermal conductivity. The relationship between the 2 properties was established by fixing the temperature and expressing conductivity as a function of apparent viscosity. Thermal conductivity increased with viscosity and decreased with increasing temperature.

Implementation of the new multichannel X-mode edge density profile reflectometer for the ICRF antenna on ASDEX Upgrade.

A new multichannel frequency modulated continuous-wave reflectometry diagnostic has been successfully installed and commissioned on ASDEX Upgrade to measure the plasma edge electron density profile evolution in front of the Ion Cyclotron Range of Frequencies (ICRF) antenna. The design of the new three-strap ICRF antenna integrates ten pairs (sending and receiving) of microwave reflectometry antennas. The multichannel reflectometer can use three of these to measure the edge electron density profiles up to 2 × 1019 m-3, at different poloidal locations, allowing the direct study of the local plasma layers in front of the ICRF antenna. ICRF power coupling, operational effects, and poloidal variations of the plasma density profile can be consistently studied for the first time. In this work the diagnostic hardware architecture is described and the obtained density profile measurements were used to track outer radial plasma position and plasma shape.

Effects of emphasising opposition and cooperation on collective movement behaviour during football small-sided games.

Optimizing collective behaviour helps to increase performance in mutual tasks. In team sports settings, the small-sided games (SSG) have been used as key context tools to stress out the players' awareness about their in-game required behaviours. Research has mostly described these behaviours when confronting teams have the same number of players, disregarding the frequent situations of low and high inequality. This study compared the players' positioning dynamics when manipulating the number of opponents and teammates during professional and amateur football SSG. The participants played 4v3, 4v5 and 4v7 games, where one team was confronted with low-superiority, low- and high-inferiority situations, and their opponents with low-, medium- and high-cooperation situations. Positional data were used to calculate effective playing space and distances from each player to team centroid, opponent team centroid and nearest opponent. Outcomes suggested that increasing the number of opponents in professional teams resulted in moderate/large decrease in approximate entropy (ApEn) values to both distance to team and opponent team centroid (i.e., the variables present higher regularity/predictability pattern). In low-cooperation game scenarios, the ApEn in amateurs' tactical variables presented a moderate/large increase. The professional teams presented an increase in the distance to nearest opponent with the increase of the cooperation level. Increasing the number of opponents was effective to overemphasise the need to use local information in the positioning decision-making process from professionals. Conversely, amateur still rely on external informational feedback. Increasing the cooperation promoted more regularity in spatial organisation in amateurs and emphasise their players' local perceptions.

Starch characterization in seven raw, boiled and roasted chestnuts (Castanea sativa Mill.) cultivars from Portugal.

Changes occurring in seven chestnut (Castanea sativa sp.) cultivars, caused by boiling and roasting, on starch content, cell and starch granules dimension were evaluated, and morphological changes were characterized by scanning electron microscopy. Three clear patterns of variation were detected after processing, namely: i) decrease of starch content with processing; ii) starch increase with the applied treatments; iii) increase of starch with boiling and decrease with roasting. Starch granules of raw chestnuts presented round, oval or elliptical form, external smooth surface and eccentric hilum, with rather ellipsoid-shaped growth rings. Processing resulted in modifications of the granules, with fusion of individual granules, and gelatinization taking place with the formation of elongated clusters. The present results indicate that boiling and roasting, besides changing the starch content of chestnut, causes important modifications in the starch granules, which can affect the sensory, rheological and chemical characteristics of chestnuts.

A fresh look at the use of nonparametric analysis in actimetry.

Actimetry has been used to estimate the sleep-wake cycle instead of the rest-activity rhythm. Although algorithms for assessing sleep from actimetry data exist, it is useful to analyze the rest-activity rhythm using nonparametric methods. This would then allow rest-activity rhythm stability, fragmentation and amplitude to be quantified. In addition, sleep and wakefulness efficiency can be quantified separately. These variables have been used in studies analyzing the effect of age, diseases and their respective treatments on human circadian rhythmicity. In this study, we carried out a comprehensive analysis of the main results from published articles and devised a functional model of interaction among the several components involved in generating the sleep-wake cycle. The nonparametric variables render it possible to infer the main characteristics of circadian rhythms, such as synchronization with a zeitgeber, and its amplitude and robustness.

The effect of different photoperiods in circadian rhythms of per3 knockout mice.

The aim of this study was to analyse the circadian behavioural responses of mice carrying a functional knockout of the Per3 gene (Per3(-/-)) to different light : dark (L : D) cycles. Male adult wild-type (WT) and Per3(-/-) mice were kept under 12-hour light : 12-hour dark conditions (12L : 12D) and then transferred to either a short or long photoperiod and subsequently released into total darkness. All mice were exposed to both conditions, and behavioural activity data were acquired through running wheel activity and analysed for circadian characteristics during these conditions. We observed that, during the transition from 12L : 12D to 16L : 8D, Per3(-/-) mice take approximately one additional day to synchronise to the new L : D cycle compared to WT mice. Under these long photoperiod conditions, Per3(-/-) mice were more active in the light phase. Our results suggest that Per3(-/-) mice are less sensitive to light. The data presented here provides further evidence that Per3 is involved in the suppression of behavioural activity in direct response to light.

Effect of defensive pressure on movement behaviour during an under-18 basketball game.

The aim of this study was to examine the effect of defensive pressure on movement behaviour during an under-18 basketball game. 20 international male players (age: M=16.05, SD=2.09 years old; weekly practice: M=10.9, SD=1.94 h; playing experience: M=7.1, SD=1.1 years) played two 10-min basketball quarters, using man-to-man »-court for the first 4 min (F»), man-to-man full court defence for the next 3 min (FULL), and man-to-man »-court defence for the last 3 min (S»). The positional data were captured by the Ubisense Real Time Location System and analysed with non-linear signal processing methods (approximate entropy) and repeated measures ANOVA. There were differences in the regularity values between F» and FULL in distance to the basket and to the opponents' basket. A stronger in-phase attraction in both lateral and longitudinal directions was identified; however, the centroids (i. e., the mean position from all team players) were closer and revealed higher values of irregularity in lateral displacements for all defensive systems. The individual speed displacements became more coordinated with teammates, particularly in the offensive court. Overall, this study provided evidence on how changing the level of defensive pressure promotes different collective behaviours.

Activity driven modeling of time varying networks.

Network modeling plays a critical role in identifying statistical regularities and structural principles common to many systems. The large majority of recent modeling approaches are connectivity driven. The structural patterns of the network are at the basis of the mechanisms ruling the network formation. Connectivity driven models necessarily provide a time-aggregated representation that may fail to describe the instantaneous and fluctuating dynamics of many networks. We address this challenge by defining the activity potential, a time invariant function characterizing the agents' interactions and constructing an activity driven model capable of encoding the instantaneous time description of the network dynamics. The model provides an explanation of structural features such as the presence of hubs, which simply originate from the heterogeneous activity of agents. Within this framework, highly dynamical networks can be described analytically, allowing a quantitative discussion of the biases induced by the time-aggregated representations in the analysis of dynamical processes.