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Coralline algae constitute one of the main groups of highly vulnerable calcified benthic organisms to ocean acidification. Although damaging effects of seawater acidification on the coralline algae skeleton have been widely demonstrated, the susceptibility to dissolution varies according to the Mg2+ in the calcite lattice. Even though the Southwest Atlantic Ocean exhibits the world's largest rhodolith beds, which occupies 20,902 km2, there is no information regarding the coralline algae species mineralogy in this area. Here, we provide mineralogical data of twenty-four coralline algae species, examine the similarity in taxonomic groups, spatial occurrence and the vulnerability of these algae to seawater acidification. Mineralogy revealed that coralline algae skeletons were mainly composed of high-Mg calcite (> 70%) with minor presence of aragonite (< 30%) and dolomite (< 3%). There were no similarities between the skeletal mineralogy of taxonomic groups and sampling regions. Remarkably, the mean Mg-substitution of encrusting coralline algae from the Brazilian Shelf was 46.3% higher than global average. Because of the higher mean Mg-substitution in calcite compared with worldwide coralline algae, these algae from Southwest Atlantic Ocean would be highly susceptible to dissolution caused by the expected near-future ocean acidification and will compromise CaCO3 net production across the Brazilian Shelf. [ABSTRACT FROM AUTHOR]

Cancer is a pathology characterized by increased cell progression and/or reduced programmed cell death. Melanoma shows a rapid increase in cell progression and its resistance to chemotherapy is associated with uncontrolled apoptosis and to mechanisms that increase the flow of the drug out of the cell. The objective of this study was to evaluate the effects of photodynamic therapy (PDT) on the cell proliferation and cellular alterations in B16F10 murine melanoma. For that, four experimental groups were evaluated: the control group; laser group (ʎ = 660 ηm, 40 mW, 2.4 J/cm2); photosensitizer group (solution containing methylene blue and toluidine blue 1:1-12.5 μg/mL); PDT group. The incubation time was 30 min. Fluorescence microscopy assays were performed without fixation with the DAPI, monodansylcadaverine (MDC), and dihydroethidium (DHE) probes. Cell proliferation was also determined at 24-h time. The tests were performed in triplicate and the statistical test used was ANOVA with Tukey post-test. The results demonstrate that the plasma membrane of the cells of all the experimental groups remained intact, ROS production and autophagy significantly increased (p < 0.0005 and p < 0.0071, respectively) only in the PDT group. The cell proliferation essay showed a reduction of 74.2% on the PDT group in relation to the control group. The present study demonstrated that oxidative stress promoted by photodynamic therapy may induce autophagy and consequently reduce cell proliferation in B16F10 melanoma. [ABSTRACT FROM AUTHOR]

• There are no systematic out-of-normal changes in the middle cerebral artery index, such as systolic, diastolic, or mean cerebral blood velocities in clinically healthy adults after a high- or moderate-intensity aerobic exercise session. • High-intensity interval exercise session provoked a more significant improvement in executive function/semantic fluency when compared to moderate-intensity continuous exercise. • Lactate increase positively correlates with improved cognitive performance after a high-intensity interval exercise session. This crossover study explored the acute effect of a session of high-intensity interval training (HIIT) or moderate-intensity continuous training (MICT) on middle cerebral artery (MCA) variables such as cerebral blood velocity, pulsatility index (PI) and resistivity index (RI) through transcranial Doppler (TCD), and cognitive function (CF - verbal fluency and Digit Span) in healthy young adults. Participants (26 healthy young adults, 13 women, 24 ± 3 years) underwent two different randomized exercise sessions: (1) MICT (60 % heart rate reserve, HRR) and (2) HIIT (80 % HRR). MCA velocity, PI, RI, CF, and serum lactate were measured immediately before and after the sessions. HIIT demonstrated improved executive function/semantic fluency (20 %, p = 0.019), while both MICT and HIIT increased lactate (625 %, HIIT, p < 0.001, and 238 %, MICT, p < 0.001). Other assessments remained stable, except for reduced PI (p = 0.029) and RI (p = 0.023) after MICT, with no significant difference (pre-post for HIIT-MICT). Notably, cognition improvement correlated with lactate increase in HIIT (ρ = 0.436; p < 0.001). Executive function/semantic fluency increased after HIIT relative to MICT. The findings show that there are no systematic out-of-normal changes in the cerebrovascular circulation of clinically healthy adults undergoing HIIT and MICT. [ABSTRACT FROM AUTHOR]

Approximately half of the Padina (Dictyotales, Phaeophyceae) species mineralize aragonite needles over the adaxial thallus surface, where mineral bands are interspersed with nonmineralized regions along the thallus from the apical to basal end. However, this calcification pattern and the related algal properties are not well understood. Therefore, this work was performed to elucidate a potential role of cell walls in the inhibition/induction of mineralization in the brown alga Padina gymnospora. In a comparison of specific thallus regions, differences were identified in the cellulose distribution, microfibrils arrangement and thickness, distribution and abundance of phenolic substances, and physical differences among the surfaces of the thallus (deformation, adhesion, topography, and nano-rugosity). In vitro mineralization assays indicated that phenolic substances are strong modulators of calcium carbonate crystals growth. In addition, de novo mineralization assays over cell wall surfaces that were used as templates, even without cellular activity, indicated that the cell wall remains a key factor in the induction/inhibition of mineralization. Overall, the current findings indicate a strong correlation between the physico-chemical and structural properties of the cell wall and the alternation pattern of the mineralization bands over the thallus of P. gymnospora. [ABSTRACT FROM AUTHOR]

This work addresses an important issue regarding the performance of simultaneous test procedures: the construction of multiple tests that at the same time are optimal from a statistical perspective and that also yield logically-consistent results that are easy to communicate to practitioners of statistical methods. For instance, if hypothesis A implies hypothesis B, is it possible to create optimal testing procedures that reject A whenever they reject B Unfortunately, several standard testing procedures fail in having such logical consistency. Although this has been deeply investigated under a frequentist perspective, the literature lacks analyses under a Bayesian paradigm. In this work, we contribute to the discussion by investigating three rational relationships under a Bayesian decision-theoretic standpoint: coherence, invertibility and union consonance. We characterize and illustrate through simple examples optimal Bayes tests that fulfill each of these requisites separately. We also explore how far one can go by putting these requirements together. We show that although fairly intuitive tests satisfy both coherence and invertibility, no Bayesian testing scheme meets the desiderata as a whole, strengthening the understanding that logical consistency cannot be combined with statistical optimality in general. Finally, we associate Bayesian hypothesis testing with Bayes point estimation procedures. We prove the performance of logically-consistent hypothesis testing by means of a Bayes point estimator to be optimal only under very restrictive conditions. [ABSTRACT FROM AUTHOR]

Abstract: In many cells, motility is mediated by flagellar beating. Protist parasites are capable of highly coordinated motility which contributes to their pathogenicity in mammalian hosts. Understanding the structural aspects of the flagellum may be important to the identification of novel targets for therapeutic intervention. Our group used atomic force microscopy (AFM) to examine the ultrastructure of Trypanosoma cruzi, obtaining valuable information on the organisation of the flagellar sub-structure. AFM images revealed novel flagellar components such as the presence of periodically-spaced protrusions organised along a flagellar furrow and oriented through the major flagellar axis between the axoneme and the paraflagellar rod. The nature and functional role of this structure are still unknown, although the hypothesis that the furrow might physically separate the two distinct domains of the flagellar membrane that comprise the surface of the axoneme and the paraflagellar rod (PFR) has been raised. To test whether the furrow was present or not only in PFR-bearing flagella, different protists containing or lacking the PFR, were analysed by AFM. Analysis of T. cruzi, Trypanosoma brucei and Herpetomonas megaseliae, which present distinct PFRs, showed similar and equivalent furrows along the main axis of their flagella, whereas Crithidia deanei, Giardia lamblia and Tritrichomonas foetus (in which the PFR is reduced or absent) lacked a furrow. Our results strongly suggest that the flagellar furrow is a characteristic feature of PFR-containing flagella and opens new perspectives for its functional role in the definition of sub-domains on the flagellar membrane. [Copyright &y& Elsevier]

Abstract: The flagellar pocket and the cytostome are surface domains of Trypanosoma cruzi epimastigote involved in acquisition of nutrients. The cytostome is physically connected to the flagellar complex. To investigate if this association plays a role in endocytosis in T. cruzi, the endocytic activity in wild type and gp72 null mutant (flagellum-cell body attachment region is absent) epimastigotes was compared. Both wild type and mutant cells were incubated with transferrin conjugated with Alexa 543 or gold particles over different time periods and thereafter qualitatively and quantitatively analyzed by flow cytometry and transmission electron microscopy. Flow cytometry analysis showed a reduction in transferrin uptake by null mutant after 30min of incubation. In addition, at this time period, signals detected by fluorescence microscopy were slightly lower in null mutant cells. At lower incubation times, no differences between wild type and mutant epimastigotes could be observed. Quantitative data obtained by morphometric and flow cytometry analysis suggested that the speed of the endocytic process in the null mutant was similar to wild type cells, although null mutants were not able to bind cargo and therefore internalize as much as wild type epimastigotes. Our observations suggest that the physical association between cytostome and the flagellar complex plays a role in endocytosis efficiency by epimastigotes of T. cruzi. [ABSTRACT FROM AUTHOR]

Background: Trypanosoma cruzi, the agent of Chagas disease, is a protozoan member of the Kinetoplastidae family characterized for the presence of specific and unique structures that are involved in different cell activities. One of them is the paraflagellar rod (PFR), a complex array of filaments connected to the flagellar axoneme. Although the function played by the PFR is not well established, it has been shown that silencing of the synthesis of its major proteins by either knockout of RNAi impairs and/or modifies the flagellar motility. Methodology/Principal Findings: Here, we present results obtained by atomic force microscopy (AFM) and transmission electron microscopy (TEM) of replicas of quick-frozen, freeze-fractured, deep-etched and rotary-replicated cells to obtain detailed information of the PFR structures in regions of the flagellum in straight and in bent state. The images obtained show that the PFR is not a fixed and static structure. The pattern of organization of the PFR filament network differs between regions of the flagellum in a straight state and those in a bent state. Measurements of the distances between the PFR filaments and the filaments that connect the PFR to the axoneme as well as of the angles between the intercrossed filaments supported this idea. Conclusions/Significance: Graphic computation based on the information obtained allowed the proposal of an animated model for the PFR structure during flagellar beating and provided a new way of observing PFR filaments during flagellar beating. [ABSTRACT FROM AUTHOR]

Abstract  In this study, the geographic patterns of genetic variation of three rodent species belonging to the tribe Oryzomyini were investigated using the mitochondrial cytochrome b and nuclear IRBP genes in biomes that are undergoing degradation processes to a greater or lesser degree. The samples are from 25 collecting localities distributed throughout the Amazon, Cerrado, Atlantic Forest, and Pampa biomes. The results show that the three species have a population and geographic structure, besides being in demographic equilibrium. The phylogenetic analyses performed on Euryoryzomys russatus and Hylaeamys megacephalus showed these specimens grouped in three distinct clades forming geographic gradients (North–South direction in H. megacephalus). Intraspecific genetic divergence was higher in H. megacephalus (4.53%), followed by E. russatus (1.79%), and lowest in Sooretamys angouya (0.88%). The results obtained indicate that, necessarily, the management strategies to preserve genetic diversity should be different for each species, since each of them presented specific population parameters. [ABSTRACT FROM AUTHOR]

Abstract: The flagellar attachment zone (FAZ) is an adhesion region of Trypanosoma cruzi epimastigote forms where the flagellum emerges from the flagellar pocket and remains attached to the cell body. This region shows a junctional complex which is formed by a linear series of apposed macular structures that are separated by amorphous material and clusters of intramembranous particles. Two protein groups appear to be important in the FAZ region: a membrane glycoprotein of 72kDa and several high molecular weight proteins. To gain a better understanding of the FAZ region, we compared wild-type Y strain T. cruzi epimastigotes with a mutant cell in which the 72-kDa surface glycoprotein (Gp72), involved in cell body-flagellum adhesion, had been deleted by target gene replacement. Using immunofluorescence confocal microscopy and electron microscopy techniques to analyze the FAZ region the results suggest that, in the absence of Gp72, other proteins involved in the formation of FAZ remain concentrated in the flagellar pocket region. The analysis of a 3-D reconstruction model of wild-type epimastigotes showed that the endoplasmic reticulum and mitochondrion are in intimate association with FAZ, in contrast to the null mutant cells where the endoplasmic reticulum was not visualized. [Copyright &y& Elsevier]

Background: Acute cellular rejection beyond the 6th month posttransplant is an uncommon complication after liver transplantation. The inadequate immunosuppression (IS) remains the main risk factor. We report a case of acute cellular rejection after a switch to everolimus monotherapy at 11 months following liver transplantation. Case presentation: This was a 69-year-old man who underwent liver transplantation after hepatocellular carcinoma. The initial immunosuppression was a combination of three immunosuppressive drugs (corticosteroids + tacrolimus + mycophenolate mofetil). The corticosteroid therapy was stopped at the 4th month posttransplant. Serious side effects of the immunosuppressive drugs (agranulocytosis and renal dysfunction), which occurred 4 months after transplantation, required a reduction and then a discontinuation of tacrolimus and mycophenolate mofetil. Everolimus was introduced as a replacement. The patient was consulted at 11 months after liver transplantation, 1 month after stopping the two immunosuppressive drugs, for liver function test abnormalities such as cytolysis and anicteric cholestasis. A moderate late acute cellular rejection was confirmed by a liver biopsy. A satisfactory biological evolution was observed following corticosteroid boluses and optimization of basic immunosuppressive drugs. Conclusion: Late acute cellular rejection remains an uncommon complication, observed mostly in the first year after liver transplantation. The main risk factor is usually the decrease of immunosuppression. [ABSTRACT FROM AUTHOR]

Multiple hypothesis testing, an important quantitative tool to report the results of scientific inquiries, frequently leads to contradictory conclusions. For instance, in an analysis of variance (ANOVA) setting, the same dataset can lead one to reject the equality of two means, say μ1= μ2, but at the same time to not reject the hypothesis that μ1= μ2= 0. These two conclusions violate the coherence principle introduced by Gabriel in 1969, and lead to results that are difficult to communicate, and, many times, embarrassing for practitioners of statistical methods. Although this situation is common in the daily life of statisticians, it is usually not discussed in courses of statistics. In this work, we enrich the teaching and discussion of this important topic by investigating through a few examples whether several standard test procedures are coherent or not. We also discuss the relationship between coherent tests and measures of support. Finally, we show how a Bayesian decision-theoretical framework can be used to build coherent tests. These approaches to coherence enlighten when such property is appealing in multiple testing and provide means of obtaining it. [ABSTRACT FROM PUBLISHER]

Decellularized cardiac extracellular matrix scaffolds with preserved composition and architecture can be used in tissue engineering to reproduce the complex cardiac extracellular matrix. However, evaluating the extent of cardiomyocyte repopulation of decellularized cardiac extracellular matrix scaffolds after recellularization attempts is challenging. Here, we describe a unique combination of biochemical, biomechanical, histological, and physiological parameters for quantifying recellularization efficiency of tissue-engineered cardiac patches compared with native cardiac tissue. Human embryonic stem cell-derived cardiomyocytes were seeded into rat heart atrial and ventricular decellularized cardiac extracellular matrix patches. Confocal and atomic force microscopy showed cell integration within the extracellular matrix basement membrane that was accompanied by restoration of native cardiac tissue passive mechanical properties. Multi-electrode array and immunostaining (connexin 43) were used to determine synchronous field potentials with electrical coupling. Myoglobin content (~60%) and sarcomere length measurement (>45% vs 2D culture) were used to evaluate cardiomyocyte maturation of integrated cells. The combination of these techniques allowed us to demonstrate that as cellularization efficiency improves, cardiomyocytes mature and synchronize electrical activity, and tissue mechanical/biochemical properties improve toward those of native tissue. [ABSTRACT FROM AUTHOR]

Decellularized cardiac extracellular matrix scaffolds with preserved composition and architecture can be used in tissue engineering to reproduce the complex cardiac extracellular matrix. However, evaluating the extent of cardiomyocyte repopulation of decellularized cardiac extracellular matrix scaffolds after recellularization attempts is challenging. Here, we describe a unique combination of biochemical, biomechanical, histological, and physiological parameters for quantifying recellularization efficiency of tissue-engineered cardiac patches compared with native cardiac tissue. Human embryonic stem cell-derived cardiomyocytes were seeded into rat heart atrial and ventricular decellularized cardiac extracellular matrix patches. Confocal and atomic force microscopy showed cell integration within the extracellular matrix basement membrane that was accompanied by restoration of native cardiac tissue passive mechanical properties. Multi-electrode array and immunostaining (connexin 43) were used to determine synchronous field potentials with electrical coupling. Myoglobin content (~60%) and sarcomere length measurement (>45% vs 2D culture) were used to evaluate cardiomyocyte maturation of integrated cells. The combination of these techniques allowed us to demonstrate that as cellularization efficiency improves, cardiomyocytes mature and synchronize electrical activity, and tissue mechanical/biochemical properties improve toward those of native tissue. [ABSTRACT FROM AUTHOR]

Cryptococcus neoformans is a fungal pathogen that causes life-threatening infections in immunocompromised individuals. Its main virulence factor is an extracellular polysaccharide capsule whose structure, assembly and dynamics remain poorly understood. In this study, we apply improved protocols for sample preparation and recently-developed scanning microscopy techniques to visualize the ultrastructure of the C. neoformans capsule at high-resolution (up to 1 nm) and improved structural preservation. Although most capsule structures in nature consist of linear polymers, we show here that the C. neoformans capsule is a ‘microgel-like’ structure composed of branched polysaccharides. Moreover, we imaged the capsule-to-cell wall link, which is formed by thin fibers that branch out of thicker capsule filaments, and have one end firmly embedded in the cell wall structure. Together, our findings provide compelling ultrastructural evidence for a branched and complex capsule conformation, which may have important implications for the biological activity of the capsule as a virulence factor. [ABSTRACT FROM AUTHOR]

NaLtural laminin matrices are formed on cell membranes by a cooperative process involving laminin self-polymerization and binding to cognate cellular receptors. In a cell-free system, lami- nm can self-polymerize, given that a minimal critical concentration is achieved. We have previously described that pH acidification renders self-polymerization independent of protein concentration. Here we studied the ultrastructure of acid-induced laminin polymers using electron and atomic force microscopies. Polymers presented the overall appearance of natural matrices and could be described as homogeneous polygonal sheets, presenting struts of 21 ± 5 and 86 ± 3 nm of height, which approximately correspond to the sizes of the~ short and the long arms of the molecule, respectively. The addition of fragment E3 (the distal two domains of the long arm) did not affect the polymerization in solution nor the formation of adsorbed matrices. On the other hand, the addition of fragment El', which contains two intact short arms, completely disrupted polymerization. These results indicate that acid-induced polymers, like natural ones, invo've only interactions between the short arms. The electrostatic surface map of laminin al LG4 -5 shows that acidification renders the distal end in the long arms exclusively positive, precluding homophylic interactions between them. Therefore, acidification reproduces in vitro, and at a physiological protein concentration, what receptor interaction does in the cellular context, namely, it prevents the long arm from disturbing formation of the homogeneous matrix involving the short arms only. We propose that acid-induced polymers are the best tool to study cellular response to laminin inthe future. [ABSTRACT FROM AUTHOR]

• Acid erosion causes surface changes in dentin. • Some NaF-containing materials are capable of preventing superficial dentin changes. • Materials showed increase in dentine inorganic content of Ca + and P-minerals. • Varnish containing 22600 ppm NaF showed dentin prevention from acid attack. • NaF + fTCP toothpaste showed dentin prevention from acid attack. The aim of this study was to evaluate the morphological and chemical effect of in-office and at-home desensitising agents containing sodium fluoride (NaF) on eroded root dentine in vitro. Fifty bovine dentine samples were pre-eroded and randomised into five groups (n = 10): G1 (Control) - milli-Q water; G2 - fluoride varnish containing NaF 22,500 ppm; G3 - desensitising cream containing NaF 9,000 ppm associated with 20% nanohydroxyapatite; G4 - toothpaste with NaF 5,000 ppm associated to tricalcium phosphate; G5 - toothpaste containing NaF 900 ppm and casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF). The specimens were submitted to erosive challenge for three days. The analyses were performed using non-contact profilometry for volumetric (Sa) and linear roughness (Ra) followed by scanning electron microscopy (SEM) and Energy Dispersive X-ray Spectrometry (EDS). The data were analysed by Kruskal-Wallis and Wilcoxon tests (α = 0.05). There was a significant reduction of Ra and Sa for the eroded samples from the G2 and G5 (p < 0.05) after an erosive challenge. The dentine surface topography pattern showed partially or totally occluded dentinal tubules after treatments, except in the control group. The control, G4 and G3 groups showed a reduction in the dentine inorganic content percentage of Ca (Calcium) and P (Phosphorus) minerals. The fluoride varnish and CPP-ACPF toothpaste were able to prevent morphological changes and were the only materials that showed the Ca and P content increased after treatment. These materials may be promising alternatives in the clinical control of dentin erosion. [ABSTRACT FROM AUTHOR]

Invasive fungal infections, including cryptococcosis, are a growing threat to immunocompromised patients. Although Cryptococcus neoformans and Cryptococcus gattii are the main agents of human cryptococcosis, opportunistic infections by environmental species, such as C. liquefaciens, have been observed recently. The main Cryptococcus virulence factor is the production and secretion of polysaccharides (PS). Previously, we showed that both species produce PS of similar composition. Here, we examined the ultrastructure and biological activity of capsular and secreted PS from C. liquefaciens, and yeast pathogenicity to an invertebrate host, in comparison with C. neoformans. Ultrastructural analysis by high-resolution microscopy showed that both species produce large and complex capsules. PS from both species had indistinguishable effects on phagocytosis levels, NO production and the secretion of a variety of immune mediators. Challenge with C. liquefaciens or C. neoformans led to complete lethality of G. mellonella larvae. Treatment with C. liquefaciens PS could not protect mice against infection with C. neoformans. We conclude that polysaccharides of the environmental yeast C. liquefaciens have strikingly similar ultrastructural and biological properties to those of C. neoformans, highlighting the importance of monitoring the emergence of new fungal pathogens for which thermotolerance may be an important transitional step towards pathogenesis in humans. [ABSTRACT FROM AUTHOR]