Trapping Microparticles in a Structured Dark Focus.

We experimentally demonstrate stable trapping and controlled manipulation of silica microspheres in a structured optical beam consisting of a dark focus surrounded by light in all directions-the dark focus tweezer. Results from power spectrum and potential analysis demonstrate the nonharmonicity of the trapping potential landscape, which is reconstructed from experimental data in agreement to Lorentz-Mie numerical simulations. Applications of the dark tweezer in levitated optomechanics and biophysics are discussed.

Antinociception induced by artemisinin nanocapsule in a model of postoperative pain via spinal TLR4 inhibition.

Artemisinin (ART) was initially described for the control of inflammation and pain. However, the mechanisms involved with its antinociceptive effect are still poorly understood. Thus, this present study aimed to investigate the effect of ART in both free and nanocapsulated form on postoperative pain, as well as the participation of the spinal Toll-like receptor 4 (TLR4) in this process. Postoperative pain was induced using the skin/muscle incision retraction (SMIR) model in male Swiss mice. After 3 and 28 days of SMIR, the animals received an intrathecal injection of free or nanocapsulated ART, and the nociceptive threshold was evaluated by von Frey filament test. To evaluate the involvement of the microglia, astrocytes, and TLR4, minocycline (a microglia inhibitor), fluorocitrate (an astrocyte inhibitor), and Lipopolysaccharide Rhodobacter sphaeroides (LPS-RS), a TLR4 antagonist, were intrathecally injected on the third day of SMIR. The levels of spinal TLR4 protein and proinflammatory cytokines tumor necrosis factor-alpha (TNF-α), and interleukin-1-beta (IL-1ß) were quantified by western blot and enzyme-linked immunosorbent assay, respectively. The results showed that free ART reduced postoperative pain (P < 0.001, F5,30 = 7.49, 16.66% for 1000 ng dose; and P < 0.01, F5,30 = 7.49, 14.58% for 500 ng dose) on the 3rd day of SMIR; while the ART nanocapsule had this effect on both the third (P < 0.001; F5,30 = 4.94; 43.75, 39.58 and 72.91% for the 250, 500 and 1000 ng doses, respectively) and 28th (P < 0.05; F5,30 = 7.71; 29.16 and 33.33% for the 500 and 1000 ng doses, respectively) day. The ART nanocapsule had a more potent and longer antinociceptive effect than free ART or morphine. Postoperative pain was also reduced by minocycline and LPS-RS. The ART nanocapsule also reduced the increased levels of TLR4, TNF-α, and IL-1ß induced by SMIR. These data suggest that the ART nanocapsule has a potent analgesic effect on postoperative pain at the spinal level, and this response involves the inhibition of TLR4 and the proinflammatory cytokines TNF-α and IL-1ß.

Essential oils as antibacterial agents against food-borne pathogens: Are they really as useful as they are claimed to be?

Most studies evaluating the use of essential oils (EO) as antibacterial agents focus mainly on minimal inhibitory concentrations (MIC) rather than minimal bactericidal concentrations (MBC). In this work, we compared MICs and MBCs of EO from condiment plants commonly used in Mediterranean Europe, namely Origanum vulgare, Salvia lavandulaefolia, Salvia officinalis, Salvia sclarea and Rosmarinus officinalis, aiming to evaluate their application as disinfecting agents in minimally processed produce. Outbreaks-related pathogens such as Listeria monocytogenes, Pseudomonas aeruginosa and Yarrowia lipolytica were used. Results showed that all EO were able to reduce bacterial growth in all bacterial strains tested, particularly O. vulgare. However, fewer EO exhibited bactericidal activities, and were only effective against one or two bacterial strains, hence eliminating the possibility to use them as broad range disinfectants. Furthermore, the necessary concentrations were too high for food application. Hence, our work suggests the need to evaluate MBC rather than MIC and questions EO usefulness in controlling undesired microorganisms. Overall, and despite the large volume of data published on EO, results obtained were not very encouraging for a realistic application on produce and question the viability of EOs as disinfecting agents in food.

Genetic variability of CYP3A4 in a heterogeneous Brazilian population from Maranhão.

Inter-individual variability in drug metabolism may result in adverse drug responses. Pharmacogenetic studies have shown that polymorphisms in drug metabolizing enzymes may contribute to this variability. Among these enzymes, CYP3A4 is responsible for metabolizing over 50% of the clinically used drugs. The Brazilian population is composed of people with Native American, European, and African ancestries, and is therefore considered as one of the most intermixed populations in the world. A thorough knowledge of the genetic frequencies of CYP3A4 allelic variants is useful for the establishment of better pharmacological therapies; therefore, the aim of this study was to describe the polymorphic frequencies for CYP3A4 -392A>G (rs2740574) in a sample population from Maranhão, Brazil. Our results showed that 75.1, 21.9, and 3.0% of the individuals expressed the -392AA, -392AG, and -392GG genotypes, respectively. The -392A and -392G alleles were observed in 86.1 and 13.9% of the population, respectively. Our results reiterate the need for a better understanding of the variations in the genotype and allele frequencies of CYP3A4 -392A>G polymorphisms in various Brazilian regions, in order to elucidate the variability in drug response.

Identification and evaluation of polymorphisms in FABP3 and FABP4 in beef cattle.

Single nucleotide polymorphisms (SNPs) were screened in FABP3 and FABP4 by automatic sequencing of pools of DNA from crossbred animals whose phenotypes belonged to the upper and lower extremes for back fat and marbling, as well as of a pool of DNA from sires used for crossbreeding. Five SNPs were identified in FABP3 and another nine SNPs were identified in FAPB4. Of these, only one SNP had no previous registry in the SNAP database (dbSNP). Three polymorphisms were selected for further evaluation of their association with production traits using restriction fragment length polymorphism-PCR (RFLP-PCR) or real-time PCR genotyping. All 3 markers were in Hardy-Weinberg equilibrium at the 5% significance level for all 7 genetic groups analyzed. Significant association was observed between FABP3-G/A with rib eye area (P = 0.035) and the rib eye area/hot carcass weight ratio (P = 0.025) and between FABP4/TasI with marbling (P = 0.052) and meat texture (P = 0.053). No significant association was observed between the FABP4-G/C polymorphism and any of the observed traits. Previous association studies with allelic variants in these genes have shown mixed results, probably because of the small effect of the genes for these traits, which suggests that results should be replicated in other populations.

Reproducibility of hypocapnic cerebrovascular reactivity measurements using BOLD fMRI in combination with a paced deep breathing task.

It has recently been proposed that hypocapnic cerebrovascular reactivity (CVR) can be assessed by measuring the blood oxygenation level dependent (BOLD) response to paced deep breathing (PDB) tasks inducing mild hypocapnia and vasoconstriction. In this work, we aim to assess the test-retest reproducibility and inter-subject variability of BOLD CVR measurements obtained using a PDB task and different methods to analyse the associated BOLD signal. The respiratory protocol consisted of alternating 40s of PDB with normal free breathing; expired CO2 pressure levels (PETCO2) were continuously monitored. CVR was quantified using either a timecourse curve analysis (TCA) approach, where the magnitude of response peaks is emphasized, or general linear modelling (GLM) including optimisation of the BOLD response latencies. The GLM fit was carried out using two types of response regressors: one that was computed as the convolution of PETCO2 traces with a gamma function and another that consisted of the convolution of PDB paradigm blocks with a physiological model of the respiratory response. Haemodynamic response latencies were optimised either on a voxel basis or for the whole imaging region. We found that the GLM method based on PDB task or PETCO2 traces and voxelwise optimisation of response latencies provided the most reproducible measures of CVR. For the average grey matter CVR, the inter-subject coefficient of variation (CVinter) / intra-subject coefficient of variation (CVintra) / intra-class correlation coefficient (ICC) were 20%/8%/0.8 and 27%/8%/0.9, using the task and PETCO2 timecourses, respectively. In terms of the spatial reproducibility, the group mean (±standard deviation) of the spatial ICC (ICCspatial) was 1.04±0.23 and 1.02±0.26, for the task and PETCO2 timecourses, respectively. These results indicate generally good reproducibility of the hypocapnic CVR maps obtained using the proposed PDB task and analysis methodology. This suggests that such protocol may therefore offer a promising alternative to conventional vasoactive challenges, which avoids their discomfort and difficulty.

Ligninolytic enzyme potential of Trametes spp. associated with leaf litter in riparian forest of the Amazônia region.

The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.

Protein stabilization with a dipeptide-mimic triazine-scaffolded synthetic affinity ligand.

Protein stabilization was achieved by a novel approach based on the adsorption and establishment of affinity-like interactions with a biomimetic triazine-scaffolded ligand. A synthetic lead compound (ligand 3'/11, K(a) ≈ 10(4) M(-1)) was selected from a previously screened solid-phase library of affinity ligands for studies of adsorption and stabilization of cutinase from Fusarium solani pisi used as a model system. This ligand, directly synthesized in agarose by a well-established solid-phase synthesis method, was able to strongly bind cutinase and led to impressive half-lives of more than 8 h at 70 °C, and of approximately 34 h at 60 °C for bound protein (a 25- and 57-fold increase as compared with the free enzyme, respectively). The ligand density in the solid matrix was found to be a determinant parameter for cutinase stabilization. It is conceivable that the highly stabilizing effect observed results from the binding of more than one ligand residue to the enzyme, creating specific macromolecular configurations that lock structural mobility thus improving molecular stability.

Pterodon pubescens oil: characterisation, certification of origin and quality control via mass spectrometry fingerprinting analysis.

INTRODUCTION: The oil obtained from Pterodon pubescens (Leguminosae) seeds are known to display anti-cancer, anti-dermatogenic and anti-nociceptive activitiy. Phytochemical studies have demonstrated that its main constituents are diterpenoids with voucapan skeletons. Considering the potential biological activities of the oil, rapid and efficient methods for assessing its quality would facilitate certification and quality control. OBJECTIVE: To develop a direct mass spectrometric fingerprinting method for the P. pubescens seed oil that would focus on the major diterpenoids constituents, enabling quality control, origin certification and recognition of marker species in commercially available products. METHOD: Two techniques were used: (i) direct infusion electrospray ionisation (ESI) mass spectrometry after solvent extraction and dilution and (ii) ambient desorption/ionisation via easy ambient sonic-spray ionisation, EASI(+)-MS, performed directly on the seed surface or at a paper surface imprinted with the oil. RESULTS: From a combination of ESI-MS, HRESI-MS and ESI-MS/MS data, 12 diterpenes were characterised, and typical profiles were obtained for the oil extract or the crude oil via both ESI-MS and EASI-MS. These techniques require no or very simple sample preparation protocols and the whole analytical processes with spectra acquisition take just a few minutes. CONCLUSION: Both techniques, but particularly EASI-MS, provide simple, fast and efficient MS fingerprinting methodologies to characterise the P. pubescens oil with typical (di)terpene profiles being applicable to quality control and certification of authenticity and origin.

The importance, prevalence and determination of vitamins B6 and B12 in food matrices: A review.

Vitamins are a vast group of fundamental organic compounds, which are not produced by the human body but are essential for the living organisms' good health. Vitamins B6 and B12 belong to the same group of hydrophilic vitamins. Structurally unrelated, they share the same purpose as essential components for normal cellular operation, growth and development. Vitamin B6 is an enzymatic co-factor that is vital for countless biochemical reactions, and is also important in sugar and fatty acid metabolization. It encompasses three natural and inter-convertible pyridine-derivatives: pyridoxine, pyridoxal and pyridoxamine. Vitamin B12 is a cobalt organometallic complex also indispensable in numerous human physiological functions. It has four bioactive forms: cyanocobalamin, methylcobalamin, hydroxocobalamin and 5'-deoxyadenosylcobalamin, and only a few prokaryotes have the ability to biosynthesize cobalamin. This work reviews the significant aspects of vitamins B6 and B12: their vital roles, consequences of deficit; food sources; and methods of determination and respective matrices, with heavy emphasis on chromatographic techniques developed within the last two decades.

Comparison of pericapsular nerve group (PENG) block and epidural analgesia following total hip arthroplasty: A retrospective analysis.

INTRODUCTION AND OBJECTIVES: Total hip arthroplasty (THA) is an increasingly common orthopaedic procedure, with moderate to severe postoperative pain. Pericapsular nerve group (PENG) block is a recent block that seems to provide adequate analgesia without significant motor blockade. The aim of this study is to retrospectively compare the analgesic efficacy and safety of PENG block with those of epidural analgesia, in patients undergoing THA. MATERIAL AND METHODS: This is a retrospective observational study of patients who underwent primary THA, submitted to epidural analgesia or single-shot ultrasound-guided PENG block, during a one-year period. Data regarding demographic characteristics, surgery and anaesthesia techniques, pain scores, opioid consumption, complications and time to hospital discharge were retrieved from institutional records and compared between the 2 groups (epidural analgesia vs PENG block). RESULTS: No significant difference was found regarding pain scores, opioid consumption, and mean time to hospital discharge between the 2 groups. Pain scores at rest (1.20 epidural vs 1.67 PENG) or with movement (3.95 epidural vs 3.72 PENG) were similar between groups. Total number of complications was higher in the epidural analgesia group (50 % epidural vs 5% PENG). Paresthesia was reported in both groups. Motor block, sedation, nausea and catheter-related complications were only found in the epidural analgesia group. CONCLUSIONS: PENG block seems to be equivalent to epidural analgesia regarding quality of postoperative analgesia for patients subject to primary THA, supporting routine use of this block in these patients. The low rate of reported complications limits conclusions on this topic.

High-density SNP association study and copy number variation analysis of the AUTS1 and AUTS5 loci implicate the IMMP2L-DOCK4 gene region in autism susceptibility.

Autism spectrum disorders are a group of highly heritable neurodevelopmental disorders with a complex genetic etiology. The International Molecular Genetic Study of Autism Consortium previously identified linkage loci on chromosomes 7 and 2, termed AUTS1 and AUTS5, respectively. In this study, we performed a high-density association analysis in AUTS1 and AUTS5, testing more than 3000 single nucleotide polymorphisms (SNPs) in all known genes in each region, as well as SNPs in non-genic highly conserved sequences. SNP genotype data were also used to investigate copy number variation within these regions. The study sample consisted of 127 and 126 families, showing linkage to the AUTS1 and AUTS5 regions, respectively, and 188 gender-matched controls. Further investigation of the strongest association results was conducted in an independent European family sample containing 390 affected individuals. Association and copy number variant analysis highlighted several genes that warrant further investigation, including IMMP2L and DOCK4 on chromosome 7. Evidence for the involvement of DOCK4 in autism susceptibility was supported by independent replication of association at rs2217262 and the finding of a deletion segregating in a sib-pair family.

Ombrohydrochory in Thismia panamensis (Standley) Jonk: a mycoheterotrophic species in Brazilian Cerrado forests.

Species of the genus Thismia Griff. are small herbs, considered mycoheterotrophic due to an intimate relationship with fungi. They are achlorophyllous, with complex floral structure but little information on reproductive strategies. This study evaluated structural and ecological aspects associated with the dispersal of seeds of Thismia panamensis (Standley) Jonk. The study was carried out in a forest fragment in the Brazilian Cerrado. During the reproductive period, 36 individuals were monitored for spatial distribution of the population and their fruits were collected. Samples were subjected to light microscopy and microtomography techniques, in addition to an experiment to evaluate seed dispersal by water droplets. Thismia panamensis is up to 8-cm tall, with a tuberous root and stem, without leaves. Its fruit is dehiscent, cup-shaped, 5 ± 1 mm in diameter, containing 219.33 ± 106.70 seeds, with an average length of 0.55 ± 0.07 mm. The seeds are exposed, and their coat has a thin and lignified wall. Accumulation of secretions was observed inside the fruits. The innermost cell layer of the ovary showed typical characteristics of aquiferous parenchyma. Water splash experiments showed that the seeds reached an average distance of 44.04 ± 26.58 cm. Each splash contained, on average, 1.50 ± 1.23 seeds, with 75% of the splashes containing a single seed. A total of 239 seeds were counted in the 163 splashes evaluated. The data show potential seed dispersal by ombrohydrochory in T. panamensis, favouring its maintenance in the study area and reflecting its clumped spatial distribution.

Flexible, dense and porous chitosan and alginate membranes containing the standardized extract of Arrabidaea chica Verlot for the treatment of skin lesions.

The combination of chitosan (C) with alginate (A) has been explored for the production of dressings due to the positive results on wound healing. CA films can show a dense or porous flexible structure, with characteristics tunable for different applications. Porosity and flexibility can be achieved, respectively, by the addition of surfactants such as Kolliphor® P188 (P) and silicone-based compounds as Silpuran® 2130 A/B (S). Furthermore, composite matrices of these polysaccharides have potential applications as devices for releasing bioactive compounds to skin lesions. The purpose of this study was to evaluate the physicochemical and biological characteristics of flexible dense and porous CA membranes incorporating the standardized extract of Arrabidaea chica Verlot (A. chica), and also to analyze the release mechanism of the extract from different membrane formulations. The results show that the inclusion of P in the formulation allows obtaining porous matrices, promotes greater homogeneity of the mixture of the silicone gel with the suspension of polysaccharides, and increases the swelling of the polymer matrix. All formulations presented high stability, reaching a maximum mass loss of 18% after seven days. The formulations with S showed the best performance in terms of flexibility and strain at break. The presence of A. chica standardized extract did not affect negatively the characteristics of the membranes. Incorporation efficiencies of the bioactive compound above 87% were achieved, and the addition of P and S to the membrane formulation changed the release of the A. chica extract kinetics. In addition, the developed formulations did not significantly affect Vero cells proliferation.

Correction to: α-Phellandrene attenuates tissular damage, oxidative stress, and TNF-α levels on acute model ifosfamide-induced hemorrhagic cystitis in mice.

The published online version contains figure in poor quality.

α-Phellandrene attenuates tissular damage, oxidative stress, and TNF-α levels on acute model ifosfamide-induced hemorrhagic cystitis in mice.

Hemorrhagic cystitis (HC) is the major dose-limiting adverse effect of the clinical use ifosfamide (IFOS). The incidence of this side effect can be as high as 75%. Mesna has been used to reduce the risk of HC, although 5% of patients who get IFOS treatment may still suffer from HC. In previous studies, our group demonstrated that α-phellandrene (α-PHE) possesses anti-inflammatory activity, which opens the door for its study in the attenuation of HC. The objective of this study was to investigate the potential uroprotective effect of the α-PHE in the mouse model of IFOS-induced HC. In order to analyze the reduction of the urothelial damage, the bladder wet weight, hemoglobin content, and the Evans blue dye extravasation from the bladder matrix were evaluated. To investigate the involvement of neutrophil migration and lipid peroxidation and involvement of enzymatic and endogenous non-enzymatic antioxidants, the tissue markers myeloperoxidase (MPO), malondialdehyde, nitrite/nitrate (NOx), superoxide dismutase (SOD), and reduced glutathione (GSH) were evaluated. TNF-α and IL-1ß were measured by ELISA immunoassay technique. The results show that pretreatment with α-PHE significantly reduced urothelial damage that was accompanied by a decrease in the activity of MPO, MDA, and NOx levels and prevention of the depletion of SOD and GSH in bladder tissues. In the assessment of cytokines, α-PHE was able to significantly reduce TNF-α level. However, it does not affect the activities of IL-1ß. These data confirm that α-PHE exerts potent anti-inflammatory properties and demonstrates that α-PHE represents a promising therapeutic option for this pathological condition.

Prevalence of children exposed to secondhand smoke at home and in the car in Azores (Portugal).

OBJECTIVES: To identify the prevalence of exposure to secondhand smoke (SHS) at home and in the car among children attending the 4th grade in Azores. METHODS: This is a cross-sectional study that assessed children's exposure to SHS in a convenience sample of school children attending the 4th grade in all primary schools of Azores. The entire population of 4th graders from all elementary schools in Azores were asked to participate in the study (n=2463) in 2017. A validated self-reported questionnaire was administered to 2092 students who delivered the signed informed consent form. We analyzed frequencies, contingency tables, and performed Chi-square tests. RESULTS: Results showed that 56.1% (95% CI 54.0-58.2) children reported having, at least, one smoking parent. Overall exposure to SHS at home was 38.4% (95% CI 36.3-40.6), and overall exposure to SHS in the car was 27.6% (95% CI 25.8-29.3). Children whose parents were smokers reported being more exposed to SHS at home (63.6%; 95% CI 58.6-68.3) than children whose parents were non-smokers (32.3%; 95% CI 30.2-34.6). CONCLUSION: The results showed that the children's exposure to SHS in Azores is high and it tends to be higher than the prevalence found in mainland Portugal. Having parents who smoke is a major risk factor for children's exposure to SHS at home. These data justify a population-wide intervention plan for preventing tobacco consumption and children's exposure to SHS in Azores.

Hemorrhagic adenoma mimicking anterior communicating artery aneurysm. / Adenoma hemorrágico que imita a un aneurisma de la arteria comunicante anterior.

TITLE: Adenoma hemorrágico que imita a un aneurisma de la arteria comunicante anterior.

Echovirus 30 detection in an outbreak of acute myalgia and rhabdomyolysis, Brazil 2016-2017.

OBJECTIVES: To describe an outbreak of acute myalgia accompanied by elevated levels of muscle enzymes that occurred in the northeast region of Brazil from December 2016 through to May 2017. METHODS: Clinical data were analysed and laboratory tests were performed in 86 specimens obtained from 52 individuals with suspected acute myalgia. A broader reactive enterovirus real-time RT-PCR followed by a semi-nested PCR amplification of partial VP1 gene were performed to identify the causative agent. RESULTS: Eighty-six clinical samples were received in our laboratory during the myalgia outbreak. Median age of individuals was 39 years. Sudden acute myalgia and dark urine were the most common symptoms. Creatine phosphokinase levels were elevated with mean value ∼16 893 U/L. Human enterovirus was detected in 67% (58/86) of the patient's specimens (urine, serum, faeces and rectal swab). The enterovirus positivity per patient was 82.7% (43/52). Echovirus 30 (E-30) (82% of the typed specimens, 18/22; 76.4% (13/17) of the typed specimens per patient) was the main enterovirus identified. In addition to E-30, CV-A16 (1/22) and E-6 (3/22) were detected in 4% and 14% of the typed specimens, respectively. No deaths occurred. CONCLUSION: The 2016-2017 outbreak of acute myalgia that occurred in the northeast region of Brazil can be associated with E-30. Despite the clinical manifestations, a favourable outcome was observed for all patients.