The parabasal filaments of Trichomonas vaginalis: A new filament and observations using 0.8 nm-resolution scanning electron microscopy.

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common nonviral sexually transmitted infection worldwide, with an estimated 260 million new cases annually. T. vaginalis contains organelles common to all eukaryotic cells, uncommon cell structures such as hydrogenosomes, and a complex and elaborate cytoskeleton constituting the mastigont system. The mastigont system is mainly formed by several proteinaceous structures associated with basal bodies, the pelta-axostylar complex made of microtubules, and striated filaments named the costa and the parabasal filaments (PFs). Although the structural organization of trichomonad cytoskeletons has been analyzed using several techniques, observation using a new generation of scanning electron microscopes with a resolution exceeding 1 nm has allowed more detailed visualization of the three-dimensional organization of the mastigont system. In this study, we have investigated the cytoskeleton of T. vaginalis using a diverse range of scanning probe microscopy techniques, which were complemented by electron tomography and Fast-Fourier methods. This multi-modal approach has allowed us to characterize an unknown parabasal filament and reveal the ultrastructure of other striated fibers that have not been published before. Here, we show the differences in origin, striation pattern, size, localization, and additional details of the PFs, thus improving the knowledge of the cell biology of this parasite.

In-vitro effect of a single application of CPP-ACP pastes and different fluoridated solutions on the prevention of dental caries around orthodontic brackets.

OBJECTIVE: To assess the in-vitro effect of single applications of CPP-ACP pastes and different fluoridated solutions on the prevention of dental caries around orthodontic brackets. MATERIAL AND METHODS: Tooth/bracket sets (n=65) were immersed in artificial saliva (1h at 37ºC) and randomly subjected to single applications (100µL; 1min) of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP emulsion), CPP-ACP with fluoride (CPP-ACPF emulsion), solutions of titanium tetrafluoride (TiF4) or sodium fluoride (NaF), or no treatment (CG). Multispecies biofilm (5 x 105 CFU/mL) was formed in the presence of 2% sucrose. After 24 h, the pH and the concentration of total soluble fluoride (TSF) were analyzed by culture medium. The presence of active white spot lesions (WSL) evaluated by macroscopic examination and the percent surface mineral loss (%SML) were analyzed. Also, the topography of enamel was detected by analysis of scanning electron microscopy (SEM). The data was assessed by chi-square, Kruskal-Wallis, and Mann-Whitney tests (p < 0.05). RESULTS: Fluoride-containing compounds led to a smaller pH reduction than did CPP-ACP and CG (p<0.05). There was difference in TSF between the groups (p<0.05), denoted as TiF4> NaF > CPP-ACPF > CPP-ACP > CG. Regarding the presence of WSL and %SML, the NaF group obtained lower values (p<0.05), while TiF4 and CPP-ACPF were similar (p>0.05). SEM demonstrated that fluoride-free groups had a larger surface dissolution. CONCLUSION: Fluoridated groups including solutions and CPP-ACPF were more effective than CPP-ACP in reducing enamel demineralization around orthodontic brackets after a single application.

Implications of Flagellar Attachment Zone Proteins TcGP72 and TcFLA-1BP in Morphology, Proliferation, and Intracellular Dynamics in Trypanosoma cruzi.

The highly adaptable parasite Trypanosoma cruzi undergoes complex developmental stages to exploit host organisms effectively. Each stage involves the expression of specific proteins and precise intracellular structural organization. These morphological changes depend on key structures that control intracellular components' growth and redistribution. In trypanosomatids, the flagellar attachment zone (FAZ) connects the flagellum to the cell body and plays a pivotal role in cell expansion and structural rearrangement. While FAZ proteins are well-studied in other trypanosomatids, there is limited knowledge about specific components, organization, and function in T. cruzi. This study employed the CRISPR/Cas9 system to label endogenous genes and conduct deletions to characterize FAZ-specific proteins during epimastigote cell division and metacyclogenesis. In T. cruzi, these proteins exhibited distinct organization compared to their counterparts in T. brucei. TcGP72 is anchored to the flagellar membrane, while TcFLA-1BP is anchored to the membrane lining the cell body. We identified unique features in the organization and function of the FAZ in T. cruzi compared to other trypanosomatids. Deleting these proteins had varying effects on intracellular structures, cytokinesis, and metacyclogenesis. This study reveals specific variations that directly impact the success of cell division and differentiation of this parasite.

In-vitro effect of a single application of CPP-ACP pastes and different fluoridated solutions on the prevention of dental caries around orthodontic brackets

ABSTRACT Objective: To assess the in-vitro effect of single applications of CPP-ACP pastes and different fluoridated solutions on the prevention of dental caries around orthodontic brackets. Material and Methods: Tooth/bracket sets (n=65) were immersed in artificial saliva (1h at 37ºC) and randomly subjected to single applications (100µL; 1min) of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP emulsion), CPP-ACP with fluoride (CPP-ACPF emulsion), solutions of titanium tetrafluoride (TiF4) or sodium fluoride (NaF), or no treatment (CG). Multispecies biofilm (5 x 105 CFU/mL) was formed in the presence of 2% sucrose. After 24 h, the pH and the concentration of total soluble fluoride (TSF) were analyzed by culture medium. The presence of active white spot lesions (WSL) evaluated by macroscopic examination and the percent surface mineral loss (%SML) were analyzed. Also, the topography of enamel was detected by analysis of scanning electron microscopy (SEM). The data was assessed by chi-square, Kruskal-Wallis, and Mann-Whitney tests (p < 0.05). Results: Fluoride-containing compounds led to a smaller pH reduction than did CPP-ACP and CG (p<0.05). There was difference in TSF between the groups (p<0.05), denoted as TiF4> NaF > CPP-ACPF > CPP-ACP > CG. Regarding the presence of WSL and %SML, the NaF group obtained lower values (p<0.05), while TiF4 and CPP-ACPF were similar (p>0.05). SEM demonstrated that fluoride-free groups had a larger surface dissolution. Conclusion: Fluoridated groups including solutions and CPP-ACPF were more effective than CPP-ACP in reducing enamel demineralization around orthodontic brackets after a single application.

RESUMO Objetivo: Avaliar in-vitro o efeito de uma aplicação única de cremes dentais de CPP-ACP e diferentes soluções fluoretadas na prevenção da cárie dentária ao redor de braquetes ortodônticos Material e Métodos: O conjunto dentes/braquetes (n=65) foi imerso em saliva artificial (1h em 37°C) randomizado e submetido a tratamento único (100µL; 1 min) de emulsão de fosfopeptídeo de caseína-fosfato de cálcio amorfo (CPP-ACP) e CPP-ACP associado ao flúor (CPP-ACPF); soluções de tetrafluoreto de titânio (TiF4) e fluoreto de sódio (NaF); e ausência de tratamento (GC). Biofilmes multiespécie (5 x 105 CFU/mL) foram formados na presença de sacarose a 2%. Após 24h, o pH e a concentração de fluoreto solúvel total (FST) foram analisados pelo meio de cultura. Foram avaliadas a presença de lesões de mancha branca (LMB), por meio da análise de macroscopia visual, e a porcentagem de perda de dureza (%PD). Também foi verificada a topografia do esmalte, usando microscopia eletrônica de varredura (MEV). Os dados foram analisados pelos testes Qui-quadrado, Kruskal-Wallis e Mann-Whitney (p < 0,05). Resultados: Os compostos contendo flúor levaram a uma redução do pH menor do que o CPP-ACP e GC (p<0,05). Houve diferença no FST entre os grupos (p <0,05), sendo TiF4> NaF > CPP-ACPF > CPP-ACP > GC. Quanto à presença de LMB e à %PD, o grupo NaF obteve os menores valores (p<0,05), enquanto TiF4 e CPP-ACPF foram semelhantes (p> 0,05). A MEV demonstrou que os grupos sem flúor tiveram uma dissolução superficial maior. Conclusão: Os grupos fluoretados, incluindo soluções e CPP-ACPF, foram mais eficazes do que o CPP-ACP sem flúor na redução da desmineralização do esmalte ao redor dos braquetes ortodônticos após uma única aplicação.

Global assessment of coralline algae mineralogy points to high vulnerability of Southwestern Atlantic reefs and rhodolith beds to ocean acidification.

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.

Tissue-engineered human embryonic stem cell-containing cardiac patches: evaluating recellularization of decellularized matrix.

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.

The use of a new calcium mesoporous silica nanoparticle versus calcium and/or fluoride products in reducing the progression of dental erosion.

Objective There is increasingly common the consumption more times a day of foods and acidic drinks in the diet of the population. The present study aimed to evaluate and compare the effects of a calcium mesoporous silica nanoparticle single application of other calcium and/or fluoride products in reducing the progression of dental erosion. Methodology Half of the eroded area was covered of 60 blocks of enamel, after which the block was submitted to the following treatments: (Ca2+-MSN), casein phosphopeptide-amorphous calcium phosphate (CPP-ACP); CPP-ACP/F-(900 ppm F-); titanium tetrafluoride (TiF4 1%) (positive control); sodium fluoride (NaF 1.36%) (positive control); and Milli-Q® water (negative control) before being submitted to a second erosive challenge. A surface analysis was performed via a three-dimensional (3D) noncontact optical profilometry to assess the volumetric roughness (Sa) and tooth structure loss (TSL) and and through scanning electron microscopy (MEV). An analysis of variance (ANOVA) and Tukey's test were performed. Results Regarding Sa, all experimental groups exhibited less roughness than the control (p<0.05). The TSL analysis revealed that the Ca2+-MSN and NaF groups were similar (p>0.05) and more effective in minimizing tooth loss compared with the other groups (p<0.05). Conclusions The Ca2+-MSN and NaF treatments were superior compared with the others and the negative control.

Effect of different home-bleaching techniques for a regular or an extended time on enamel properties.

CONTEXT: The side effects of bleaching products are still incompletely known. AIMS: This work aims to evaluate the effects of bleaching regimens on colour variation, microstructure, roughness, composition and nanohardness of human dental enamel until 8 weeks. SETTINGS AND DESIGN: : Twenty teeth were cross-sectioned to obtain eighty enamel fragments (50 × 50 mm) divided into four groups: CN (control Negative-artificial saliva), CP10 (10% carbamide peroxide), HP10 (10% hydrogen peroxide), and WS (whitening strips-10% hydrogen peroxide). METHODS AND MATERIAL: Roughness (atomic force microscopy-AFM and 3-D surface scanning), morphology (confocal laser scanning microscopy-CLSM and AFM), hardness and elastic modulus (nanoindentation), and composition (Raman microspectroscopy) were analysed before the therapy and after 4 and 8 weeks. Colour measures were performed weekly. STATISTICAL ANALYSIS USED: : Two-way ANOVA with repeated measures (P < 0.05). RESULTS: Bleaching stabilizes after 3 weeks for HP10 and after 4 weeks for CP10 and WS. Roughness evaluation showed statistical difference for HP10 after 8 weeks for Sa and Sq, for HP10 and WS after 4 weeks and for CP10 after 8 weeks. The same occurred for hardness and elastic modulus. The morphological evaluation demonstrated the most significant effects after 8 weeks of treatment for HP10 and WS. Composition analysis revealed modifications in peaks related to the organic content spectra (protein) with an increase in detection after 4 weeks, followed by a decrease after 8 weeks. CONCLUSIONS: H2O2-based products caused morphological and compositional alterations on enamel.

Morphological and chemical effects of in-office and at-home desensitising agents containing sodium fluoride on eroded root dentin.

OBJECTIVE: 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. METHODS: 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). RESULTS: 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. CONCLUSION: 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.

The use of a new calcium mesoporous silica nanoparticle versus calcium and/or fluoride products in reducing the progression of dental erosion

Abstract Objective There is increasingly common the consumption more times a day of foods and acidic drinks in the diet of the population. The present study aimed to evaluate and compare the effects of a calcium mesoporous silica nanoparticle single application of other calcium and/or fluoride products in reducing the progression of dental erosion. Methodology Half of the eroded area was covered of 60 blocks of enamel, after which the block was submitted to the following treatments: (Ca2+-MSN), casein phosphopeptide-amorphous calcium phosphate (CPP-ACP); CPP-ACP/F-(900 ppm F−); titanium tetrafluoride (TiF4 1%) (positive control); sodium fluoride (NaF 1.36%) (positive control); and Milli-Q® water (negative control) before being submitted to a second erosive challenge. A surface analysis was performed via a three-dimensional (3D) noncontact optical profilometry to assess the volumetric roughness (Sa) and tooth structure loss (TSL) and and through scanning electron microscopy (MEV). An analysis of variance (ANOVA) and Tukey's test were performed. Results Regarding Sa, all experimental groups exhibited less roughness than the control (p<0.05). The TSL analysis revealed that the Ca2+-MSN and NaF groups were similar (p>0.05) and more effective in minimizing tooth loss compared with the other groups (p<0.05). Conclusions The Ca2+-MSN and NaF treatments were superior compared with the others and the negative control.

Micro-shear bond strength of composite resin to glass ionomer cement using an alternative method to build up test specimens.

CONTEXT: Despite the relevance of the sandwich technique, there are still doubts about the best adhesive strategy and surface treatment for glass ionomer cements (GICs). AIM: The aim of this study is to evaluate the best surface treatment for GIC to ensure an effective and durable adhesion to resin, through micro-shear test, using an alternative method to build up test specimens. SUBJECTS AND METHODS: Eighty GIC samples were divided into eight groups (n = 10) according to five surface treatments (none, etching, air drying, grinding, and grinding plus etching) and according to the adhesive system (conventional or self-etch). Five starch tubes were positioned on each sample, and a flowable composite was inserted generating 50 resin test bodies per group and a total of 400 tested areas. All specimens were submitted to the micro-shear test: half immediately and half after thermal cycling (10,000 cycles of 20 s each/5° and 55°C). All samples were analyzed to evaluate fracture. Representative samples were also analyzed by scanning electron microscopy and energy dispersive spectroscopy. Data were analyzed with two-way ANOVA and Tukey's honest significant difference post hoc test (P <.05). RESULTS: The bond strengths in the thermal cycled specimens were lower and showed a statistically significant difference (P = 0). The "grinding" groups showed the highest bond strength. CONCLUSIONS: The alternative method to build up test specimens was effective and easy to execute. Grinding of the GIC surface, which is not normally performed before the use of the adhesive system, represented the best option of surface treatment.

Enamel sample preparation for AFM: Influence on roughness and morphology.

Human dental enamel is organized by prisms that are structured between 3 and 6 µm in diameter. Determining the relationships between different treatments on the surface of enamel using ultrastructural analysis is the purpose of many in vitro experiments. Different sample pretreatments have been reported in the literature. Grinding and polishing are common procedures for enamel preparation. They provide a flat and standardized surface, which is imperative for the use of some techniques such as ATR-FTIR. However, for morphological analysis, SEM and AFM represent easier methods to measure and reduce the biological sample variation. Therefore, the objective of this study was to establish how different forms of enamel preparation can influence the advent of artifacts during ultrastructural observation, especially by AFM analysis. Four groups (n = 10) were tested: (a) without preparation; (b) polishing with a diamond paste; (c) grinding with decreasing granulations of silicon carbide papers; (d) grinding with polishing. Images were obtained using the Peak-Force Tapping mode. After the first images were obtained, all fragments were acid etched with 37% phosphoric acid for 30 seconds, rinsed for 60 seconds, and dried intensively. Upon grinding and polishing, the exposure of the inner enamel surfaces provided a less mineralized layer that was marked by scratches and a higher susceptibility to treatments. Moreover, using native enamel provided more valuable information on the surface and the roughness changes for clinical applications. In addition, phosphoric acid is an option for observing the prismatic arrangement after grinding and/or polishing changes the morphology. RESEARCH HIGHLIGHTS: The use of native enamel samples to investigate the effects of different treatments on surface should be preferred in research, when the technique allows it.

Oral infection of mice with Fusobacterium nucleatum results in macrophage recruitment to the dental pulp and bone resorption.

BACKGROUND: Fusobacterium nucleatum is a Gram-negative anaerobic bacterium associated with periodontal disease. Some oral bacteria, like Porphyromonas gingivalis, evade the host immune response by inhibiting inflammation. On the other hand, F. nucleatum triggers inflammasome activation and release of danger-associated molecular patterns (DAMPs) in infected gingival epithelial cells. METHODS: In this study, we characterized the pro-inflammatory response to F. nucleatum oral infection in BALB/c mice. Western blots and ELISA were used to measure cytokine and DAMP (HMGB1) levels in the oral cavity after infection. Histology and flow cytometry were used to observe recruitment of immune cells to infected tissue and pathology. RESULTS: Our results show increased expression and production of pro-inflammatory cytokines during infection. Furthermore, we observe that F. nucleatum infection leads to recruitment of macrophages in different tissues of the oral cavity. Infection also contributes to osteoclast recruitment, which could be involved in the observed bone resorption. CONCLUSIONS: Overall, our findings suggest that F. nucleatum infection rapidly induces inflammation, release of DAMPs, and macrophage infiltration in gingival tissues and suggest that osteoclasts may drive bone resorption at early stages of the inflammatory process.

Cell wall physicochemical properties determine the thallus biomineralization pattern of Padina gymnospora (Phaeophyceae).

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.

The environmental yeast Cryptococcus liquefaciens produces capsular and secreted polysaccharides with similar pathogenic properties to those of C. neoformans.

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.

Effect of prior silane application on the bond strength of a universal adhesive to a lithium disilicate ceramic.

STATEMENT OF PROBLEM: Universal adhesives combine silane and various monomers in a single bottle to make them more versatile. Their adhesive performance is unclear. PURPOSE: The purpose of this in vitro study was to assess the effects of an additional silane application before using a universal adhesive on the adhesion between a disilicate glass ceramic and a composite resin by using a microshear bond strength test (µSBS) and fracture analysis immediately and after thermocycling. MATERIAL AND METHODS: One hundred lithium disilicate glass ceramic disks were divided into 10 groups for bond strength testing according to the following 3 surface treatments: silane application (built-in universal adhesive or with additional application), adhesive (Adper Single Bond Plus [SB, 3M ESPE], Scotchbond Universal Adhesive [U, 3M ESPE], and mixed U with Dual Cure Activator [DCA, 3M ESPE]); or thermocycling (half of the specimens were thermocycled 10000 times). After surface treatment, 5 resin cylinders were bonded to each disk and submitted to a µSBS test. The failure mode was analyzed under a stereomicroscope and evaluated by scanning electron microscope and energy-dispersive x-ray spectroscopy. Data from the µSBS test were analyzed by 3-way ANOVA followed by the Tukey HSD post hoc test (α=.05). RESULTS: An additional silane application resulted in a higher µSBS result for all adhesive groups (P<.05). CONCLUSIONS: Ceramic surface treatment influenced the performance of adhesives, which may be improved with an additional silane application.

Cryptococcus neoformans capsular polysaccharides form branched and complex filamentous networks viewed by high-resolution microscopy.

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.

Silver nanoparticle production by the fungus Fusarium oxysporum: nanoparticle characterisation and analysis of antifungal activity against pathogenic yeasts.

The microbial synthesis of nanoparticles is a green chemistry approach that combines nanotechnology and microbial biotechnology. The aim of this study was to obtain silver nanoparticles (SNPs) using aqueous extract from the filamentous fungus Fusarium oxysporum as an alternative to chemical procedures and to evaluate its antifungal activity. SNPs production increased in a concentration-dependent way up to 1 mM silver nitrate until 30 days of reaction. Monodispersed and spherical SNPs were predominantly produced. After 60 days, it was possible to observe degenerated SNPs with in additional needle morphology. The SNPs showed a high antifungal activity against Candida and Cryptococcus , with minimum inhibitory concentration values ≤ 1.68 µg/mL for both genera. Morphological alterations of Cryptococcus neoformans treated with SNPs were observed such as disruption of the cell wall and cytoplasmic membrane and lost of the cytoplasm content. This work revealed that SNPs can be easily produced by F. oxysporum aqueous extracts and may be a feasible, low-cost, environmentally friendly method for generating stable and uniformly sized SNPs. Finally, we have demonstrated that these SNPs are active against pathogenic fungi, such as Candida and Cryptococcus.

Silver nanoparticle production by the fungus Fusarium oxysporum: nanoparticle characterisation and analysis of antifungal activity against pathogenic yeasts

The microbial synthesis of nanoparticles is a green chemistry approach that combines nanotechnology and microbial biotechnology. The aim of this study was to obtain silver nanoparticles (SNPs) using aqueous extract from the filamentous fungus Fusarium oxysporum as an alternative to chemical procedures and to evaluate its antifungal activity. SNPs production increased in a concentration-dependent way up to 1 mM silver nitrate until 30 days of reaction. Monodispersed and spherical SNPs were predominantly produced. After 60 days, it was possible to observe degenerated SNPs with in additional needle morphology. The SNPs showed a high antifungal activity against Candida and Cryptococcus , with minimum inhibitory concentration values ≤ 1.68 µg/mL for both genera. Morphological alterations of Cryptococcus neoformans treated with SNPs were observed such as disruption of the cell wall and cytoplasmic membrane and lost of the cytoplasm content. This work revealed that SNPs can be easily produced by F. oxysporum aqueous extracts and may be a feasible, low-cost, environmentally friendly method for generating stable and uniformly sized SNPs. Finally, we have demonstrated that these SNPs are active against pathogenic fungi, such as Candida and Cryptococcus .

Visualization of the flagellar surface of protists by atomic force microscopy.

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.