A new 3D model of L929 fibroblasts microtissues uncovers the effects of Bothrops erythromelas venom and its antivenom.

In Brazil, around 80% of snakebites are caused by snakes of the genus Bothrops. A three-dimensional culture model was standardized and used to perform treatments with Bothrops erythromelas venom (BeV) and its antivenom (AV). The MRC-5 and L929 cell lines were cultured at increasing cell densities. Morphometric parameters were evaluated through images obtained from an inverted microscope: solidity, circularity, and Feret diameter. L929 microtissues (MT) showed better morphometric data, and thus they were used for further analysis. MT viability was assessed using the acridine orange and ethidium bromide staining method, which showed viable cells in the MT on days 5, 7, and 10 of cultivation. Histochemical and histological analyses were performed, including hematoxylin/eosin staining, which showed a good structure of the spheroids. Alcian blue staining revealed the presence of acid proteoglycans. Immunohistochemical analysis with ki-67 showed different patterns of cell proliferation. The MT were also subjected to pharmacological tests using the BeV, in the presence or absence of its AV. The results showed that the venom was not cytotoxic, but it caused morphological changes. The MT showed cell detachment, losing their structure. The antivenom was able to partially prevent the venom activities.

Influence of prepartum dietary cation-anion difference and the magnitude of calcium decline at the onset of lactation on mineral metabolism and physiological responses.

The onset of lactation is characterized by substantially altered calcium (Ca) metabolism; recently, emphasis has been placed on understanding the dynamics of blood Ca in the peripartal cow in response to this change. Thus, the aim of our study was to delineate how prepartum dietary cation-anion difference (DCAD) diets and the magnitude of Ca decline at the onset of lactation altered blood Ca dynamics in the periparturient cow. Thirty-two multiparous Holstein cows were blocked by parity, previous 305-d milk yield and expected parturition date, and randomly allocated to either a positive (+120 mEq/kg) or negative (-120 mEq/kg) DCAD diet from 251 d of gestation until parturition (n = 16/diet). Immediately after parturition cows were continuously infused for 24 h with (1) an intravenous solution of 10% dextrose or (2) Ca gluconate (CaGlc) to maintain blood ionized (iCa) concentrations at ∼1.2 mM (normocalcemia) to form 4 treatment groups (n = 8/treatment). Blood was sampled every 6 h from 102 h before parturition until 96 h after parturition and every 30 min during 24 h continuous infusion. Cows fed a negative DCAD diet prepartum exhibited a less pronounced decline in blood iCa approaching parturition with lesser magnitude of decline relative to positive DCAD-fed cows. Cows fed a negative DCAD diet prepartum required lower rates of CaGlc infusion to maintain normocalcemia in the 24 h postpartum relative to positive DCAD-fed cows. Infusion of CaGlc disrupted blood Ca and P dynamics in the immediate 24 h after parturition and in the days following infusion. Collectively, these data demonstrate that prepartum negative DCAD diets facilitate a more transient hypocalcemia and improve blood Ca profiles at the onset of lactation whereas CaGlc infusion disrupts mineral metabolism.

Longitudinal association of sleep quality with physical performance measures: SABE cohort study, Brazil.

OBJECTIVES: The aim of this study was to investigate the longitudinal association of sleep with physical performance in a representative sample of non-institutionalised older adults residing in the municipality of São Paulo, Brazil. STUDY DESIGN: Prospective cohort study. METHODS: The current longitudinal study used data extracted from the Health, Well-being, and Aging Study (Estudo Saúde Bem-Estar e Envelhecimento [SABE]). The study population consisted of individuals aged ≥60 years who participated in the study in 2010 or 2015. Dependent variables included the Short Physical Performance Battery (SPPB) and gait speed. Independent variables of interest were self-reported sleep difficulty, daytime sleepiness and sleep quality. The longitudinal association between sleep variables and the outcomes was evaluated using Generalised Estimating Equations (GEE) Models adjusted for covariates. All the variables, except age, sex and schooling, were assessed at baseline and follow-up visits (2010 and 2015). RESULTS: The analyses included 2205 observations from 1559 individuals. The population mean age was 72 years in 2010 and 71 years in 2015, with a higher prevalence of women in both years. Between 2010 and 2015, there was a decline in the SPPB score and gait speed. Daytime sleepiness was negatively associated with the SPPB score [Coef.: -0.38 (95% confidence interval {CI}: -0.56, -0.21)] and gait speed [Coef.: -0.03 (95% CI: -0.05, -0.01)]. Poor sleep quality was negatively associated with the SPPB score [Coef.: -0.29 (95% CI: -0.57, -0.01)] and gait speed [Coef.: -0.03 (95% CI: -0.06, -0.00)]. CONCLUSIONS: Daytime sleepiness and poor sleep quality are associated with compromised physical performance in non-institutionalised older adults, and this association remained consistent over time.

Burden of non-communicable diseases and the achievement of the sustainable development goals in 2030 in Mercosur countries.

OBJECTIVES: The aims of this article were to analyse the burden of NCDs and their RFs in the Mercosur countries between 1990 and 2019 and to project mortality trends for 2030. STUDY DESIGN: Epidemiological study of time series. METHODS: The present study used data from the Global Burden of Disease study. The absolute number of deaths, mortality rates, disability-adjusted life years, years of life lost, years lived with disability and the burden of premature mortality by NCD attributable to the RFs were evaluated. Projections were made up to 2030. Age-standardised rates were used to draw comparisons by years and by countries. The analysis was conducted using the RStudio software. RESULTS: Between 1990 and 2019, a decrease was found in the premature mortality rates caused by NCDs in all the countries, except for Paraguay, which remained stable. When analysing premature mortality rates due to NCDs up to 2030, it was predicted that none of the countries would achieve the sustainable development goal of a one-third reduction in premature mortality by NCDs. Regarding the impacts of the RFs for NCDs, smoking, dietary risks, high blood pressure (BP) and high body mass index (BMI) were the main risks attributable to premature deaths due to NCDs. CONCLUSIONS: The results showed that mortality rates are declining in Mercosur countries; however, none of the countries are predicted to achieve the sustainable development goal of a one-third reduction in mortality due to NCDs by 2030. In addition to access to adequate treatment, progress is required in public regulation actions to reduce RFs, such as smoking, dietary risks, high BP and high BMI.

Chemical and mechanical influence of root canal irrigation on biofilm removal from lateral morphological features of simulated root canals, dentine discs and dentinal tubules.

AIM: To investigate the anti-biofilm efficacy of irrigation using a simulated root canal model, the chemical effect of irrigants against biofilms grown on dentine discs and their impact on biofilm viscoelasticity, the efficacy of the irrigants in decontaminating infected dentinal tubules and the capacity of bacteria to regrow. METHODOLOGY: Biofilm removal, viscoelastic analysis of remaining biofilms and bacterial viability were evaluated using a simulated root canal model with lateral morphological features, dentine discs and a dentinal tubule model, respectively. Experiments were conducted using a two-phase irrigation protocol. Phase 1: a modified salt solution (RISA) and sodium hypochlorite (NaOCl) were used at a low flow rate to evaluate the chemical action of the irrigants. Ultrasonic activation (US) of a chemically inert solution (buffer) was used to evaluate the mechanical efficacy of irrigation. Phase 2: a final irrigation with buffer at a high flow rate was performed for all groups. Optical coherence tomography (OCT), low load compression testing (LLCT) and confocal scanning laser microscopy analysis were used in the different models. One-way analysis of variance (anova) was performed for the OCT and LLCT analysis, whilst Kruskal-Wallis and Wilcoxon ranked tests for the dentinal tubule model. RESULTS: US and high flow rate removed significantly more biofilm from the artificial lateral canal. For biofilm removal from the artificial isthmus, no significant differences were found between the groups. Within-group analysis revealed significant differences between the steps of the experiment, with the exception of NaOCl. For the dentine discs, no significant differences regarding biofilm removal and viscoelasticity were detected. In the dentinal tubule model, NaOCl exhibited the greatest anti-biofilm efficacy. CONCLUSIONS: The mechanical effect of irrigation is important for biofilm removal. An extra high flow irrigation rate resulted in greater biofilm removal than US in the artificial isthmus. The mechanical effect of US seemed to be more effective when the surface contact biofilm-irrigant was small. After the irrigation procedures, the remaining biofilm could survive after a 5-day period. RISA and NaOCl seemed to alter post-treatment remaining biofilms.

Biofilm removal from a simulated isthmus and lateral canal during syringe irrigation at various flow rates: a combined experimental and Computational Fluid Dynamics approach.

AIM: (i) To quantify biofilm removal from a simulated isthmus and a lateral canal in an artificial root canal system during syringe irrigation with NaOCl at different concentrations and delivered at various flow rates (ii) to examine whether biofilm removal is further improved by a final high-flow-rate rinse with an inert irrigant following irrigation with NaOCl. (iii) to simulate the irrigant flow in these areas using a computer model (iv) to examine whether the irrigant velocity calculated by the computer model is correlated to biofilm removal. METHODOLOGY: Ninety-six artificial root canals with either a simulated isthmus or lateral canal were used. A dual-species in vitro biofilm was formed in these areas using a Constant Depth Film Fermenter. NaOCl at various concentrations (2, 5 and 10%) or adhesion buffer (control) was delivered for 30 s by a syringe and an open-ended needle at 0.033, 0.083, or 0.166 mL s-1 or passively deposited in the main root canal (phase 1). All specimens were subsequently rinsed for 30 s with adhesion buffer at 0.166 mL s-1 (phase 2). The biofilm was scanned by Optical Coherence Tomography to determine the percentage of the remaining biofilm. Results were analysed by two 3-way mixed-design ANOVAs (α = 0.05). A Computational Fluid Dynamics model was used to simulate the irrigant flow inside the artificial root canal system. RESULTS: The flow rate during phase 1 and additional irrigation during phase 2 had a significant effect on the percentage of the remaining biofilm in the isthmus (P = 0.004 and P < 0.001). Additional irrigation during phase 2 also affected the remaining biofilm in the lateral canal significantly (P ≤ 0.007) but only when preceded by irrigation at medium or high flow rate during phase 1. The effect of NaOCl concentration was not significant (P > 0.05). Irrigant velocity in the isthmus and lateral canal increased with increasing flow rate and it was substantially correlated to biofilm removal from those areas. CONCLUSIONS: The irrigant flow rate affected biofilm removal in vitro more than NaOCl concentration. Irrigant velocity predicted by the computer model corresponded with the pattern of biofilm removal from the simulated isthmus and lateral canal.

Effect of education and multimorbidity on mortality among older adults: findings from the health, well-being and ageing cohort study (SABE).

OBJECTIVES: This study assessed the moderating role of education on the relationship between multimorbidity and mortality among older adults in Brazil. STUDY DESIGN: This was a cohort study. METHODS: This study used data from 1768 participants of the Health, Well-Being and Ageing Cohort Study (SABE) who were assessed between 2006 and 2015. The Cox Proportional Risks Model was used to evaluate the association between multimorbidity (two or more chronic diseases) and mortality. An interaction term between education and multimorbidity was included to test the moderating role of education in this association. RESULTS: The average follow-up time was 4.5 years, with a total of 589 deaths in the period. Multimorbidity increased the risk of mortality (hazard ratio [HR] 1.55, 95% confidence interval [CI] 1.27-1.91), and this association was not moderated by education (HR 1.06, 95% CI 1.00-1.13; P value = 0.07). CONCLUSIONS: The impact of education and multimorbidity on mortality emphasises the need for an integrated approach directed towards the social determinants of health to prevent multimorbidity and its burden among older adults.

The influence of time and irrigant refreshment on biofilm removal from lateral morphological features of simulated root canals.

AIM: To evaluate the effect of irrigant refreshment and exposure time of a 2% sodium hypochlorite solution (NaOCl) on biofilm removal from simulated lateral root canal spaces using two different flow rates. METHODOLOGY: A dual-species biofilm was formed by a Constant Depth Film Fermenter (CDFF) for 96 h in plug inserts with anatomical features resembling an isthmus or lateral canal-like structures. The inserts were placed in a root canal model facing the main canal. NaOCl 2% and demineralized water (control group) were used as irrigant solutions. Both substances were applied at a flow rate of 0.05 and 0.1 mL s-1 . The samples were divided into three groups with zero, one or two refreshments in a total exposure time of 15 min. A three-way analysis of variance (anova) was performed to investigate the interaction amongst the independent variables and the effect of consecutive irrigant refreshment on percentage of biofilm removal. A Tukey post hoc test was used to evaluate the effect of each independent variable on percentage biofilm removal in the absence of statistically significant interactions. RESULTS: For the lateral canal, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time (P = 0.005). There was no significant effect in biofilm removal between the consecutive irrigant refreshments measured in the same biofilm. For the isthmus, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time; both NaOCl and a high flow rate removed significantly more biofilm when the exposure time was analysed (P = 0.018 and P = 0.029, respectively). Evaluating the effect of consecutive irrigant refreshment on the same biofilm, 2% NaOCl, 0.1 mL s-1 flow rate and one or two refreshments removed significant more biofilm (P = 0.04, 0.034 and 0.003, <0.001, respectively). CONCLUSIONS: In this model, refreshment did not improve biofilm removal from simulated lateral root canal spaces. NaOCl removed more biofilm from the lateral canal- and isthmus-like structure. A higher flow rate removed significantly more biofilm from the isthmus-like structure. There was always remaining biofilm left after the irrigation procedures.

Individuals at risk of seropositive rheumatoid arthritis: the evolving story.

The aetiology of the autoimmune disease rheumatoid arthritis (RA) involves a complex interplay between genetic and environmental factors that initiate many years before the onset of clinical symptoms. These interactions likely include both protective and susceptibility factors which together determine the risk of developing RA. More than 100 susceptibility loci have been linked to RA. The strongest association is with HLA-DRB1 alleles encoding antigen presenting molecules containing a unique sequence in the peptide-binding grove called the 'shared epitope'. Female sex, infections during childhood, lifestyle habits (e.g. smoking and diet) and distinct microbial agents, amongst many others, are interacting risk factors thought to contribute to RA pathogenesis by dysregulating the immune system in individuals with genetic susceptibility. Interestingly, patients with RA develop autoantibodies many years before the clinical onset of disease, providing strong evidence that the lack of tolerance to arthritogenic antigens is amongst the earliest events in the initiation of seropositive RA. Here, we will discuss the clinical and mechanistic evidence surrounding the role of different environmental and genetic factors in the phases leading to the production of autoantibodies and the initiation of symptomatic RA. Understanding this complexity is critical in order to develop tools to identify drivers of disease initiation and propagation and to develop preventive therapeutics.

Antimicrobial action of photodynamic therapy in root canals using LED curing light, curcumin and carbopol gel.

AIM: To evaluate the capacity of carbopol gel to maintain the intensity of a LED curing light (blueLED) along the length of prepared root canals in bovine teeth, and to assess the antimicrobial capacity of curcumin photoactivated by a LED curing light in the presence of carbopol gel. METHODOLOGY: Experiment 1: Eight straight roots of bovine incisors were standardized to a length of 15 mm, and the root canals instrumented up to a size 120 K-file. The LED curing light was irradiated inside the root canals using an aluminium collimator (1.5 mm in diameter) placed at the orifice (n = 8). Initially, the irradiation was performed in empty root canals and then repeated with the root canals filled with carbopol gel. Simple standardized photographs of the roots were taken with a digital camera in the mesial perspective during the irradiation procedure and the images analysed in OriginLab software to verify the light intensity along the length of the root. Experiment 2: Twenty dentine blocks were obtained from the cervical third of bovine incisors using a trephine bur. Biofilms were induced for 21 days on the blocks using Enterococcus faecalis (ATCC 4083) at 109 cells mL-1 . The blocks were treated according to the groups (n = 5): positive control; standard PDT (methylene blue + diode Laser); curcumin; LED curing light; and curcumin + LED curing light. After the treatment, the samples were dyed with Live/Dead BacLight Bacterial Viability solution and fluorescence images were obtained by Confocal Scanning Laser Microscopy (CSLM). Experiment 3: Thirty-two roots of bovine incisors were prepared as described in experiment 1. Their dentinal tubules were contaminated and the root canals treated according to the groups (n = 8): positive control; standard PDT; curcumin + LED curing light; curcumin + carbopol gel + LED curing light. The specimens were sectioned longitudinally and the split roots were treated with the Live/Dead dye to obtain fluorescence images by CSLM. All images were processed using BioImageL software to measure the percentage of viable bacteria and the data analysed statistically using the nonparametric Kruskal-Wallis test (α < 0.05). RESULTS: In Experiment 1, carbopol gel did not improve the intensity of LED light transmission along the root canal. In Experiment 2, a significant decrease (P < 0.05) in bacterial viability occurred in the following order: positive control < only LED curing light < only curcumin < curcumin + LED curing light = standard PDT; and in Experiment 3 positive control = curcumin + LED curing light ≤ curcumin + gel + LED curing light ≤ standard PDT. CONCLUSION: Similar disinfection effectiveness was obtained using curcumin + LED curing light and methylene blue + 660 nm LASER (standard PDT). The use of carbopol gel did not favour a greater transmission of LED light along the root canal and also resulted in less bacterial killing when used in endodontic PDT.

Intra-articular leflunomide-loaded poly(ε-caprolactone) implants to treat synovitis in rheumatoid arthritis.

Rheumatoid arthritis is an autoimmune pathology that manifests as chronic inflammatory arthropathy and synovitis. Treatment of rheumatoid arthritis is based on the administration of different types of drugs, including leflunomide, an antirheumatic drug. However, the long-term systemic use of leflunomide may be associated with adverse effects. Local therapy could be an efficient strategy to treat synovitis triggered by rheumatoid arthritis without inducing adverse effects. In this study, leflunomide-loaded poly(ε-caprolactone) implants (leflunomide PCL implants) were evaluated as local drug delivery systems capable of attenuating inflammation and angiogenesis, which represent events of synovitis. Leflunomide PCL implants were designed by hot molding technique; and they were characterized by FTIR and DSC. These analytical techniques demonstrated the chemical integrity and dispersion of drug into the polymeric chains. Then, a spectrophometric method was developed and validated to quantify the leflunomide incorporated into the PCL implants and released from them. Linearity was obtained by ordinary least squares regression method to estimate the linear regression equation. Residues were evaluated considering normality, independence and homoscedasticity. Precision was lower than 5 %, and accuracy ranged from 98 to 104.5 %. Quantitation limit was 2.0 µg mL-1. PCL implants provided controlled and sustained release of leflunomide for 30 consecutive days after inserting these systems in the subcutaneous tissue of mice. The main mechanisms of drug delivery were solubilization and diffusion from polymer. Then, a non-biocompatible sponge was inserted into the subcutaneous tissue of mice to function as a frame to develop the inflammatory and angiogenic processes. Leflunomide PCL implants were inserted in direct contact with the sponge. At 4, 7 and 10 days after-sponge implantation, the key components of inflammatory angiogenesis were measured to verify the regression of these events induced by drug. Leflunomide controlled released from polymeric implants downregulated the neutrophil and monocyte/macrophage infiltration due to the reduced expression of myeloperoxidase (MPO) and N-acetyl-ß-d-glucosaminidase (NAG), respectively. As the influx of these pro-inflammatory cells was modulated by leflunomide, the production of nitric oxide (NO), a pro-inflammatory substance, reached low concentrations in the sponge. As a consequence of the modulation of inflammation at the pathological site, the angiogenic process was downregulated, since the hemoglobin levels in the sponge were drastically reduced. The accumulation of leflunomide in the pathological site did not induce nephrotoxicity or hepatototoxicity, as confirmed by histological analyses. Finally, intra-articular leflunomide PCL implants represent a potential therapeutic alternative to treat locally the synovitis triggered by rheumatoid arthritis without inducing systemic adverse effects.

Complement Factor H as a potential atherogenic marker in chronic Chagas' disease.

We aimed to investigate the association between plasma levels of complement Factor H (FH) with cardiac involvement, inflammatory and cardiometabolic parameters in patients with chronic Chagas' disease (CD). FH plasmatic levels were determined in 80 chronic CD patients. Glycaemic index, lipidogram (high-density lipoprotein cholesterol HDL-C, low-density lipoprotein cholesterol LDL-C, triglycerides and total cholesterol) and Ultrasensitive C-Reactive Protein (uCRP) values were obtained from medical records. Height, weight, body mass index (BMI) blood pressure and left ventricular ejection fraction (LVEF) were obtained from echocardiography examinations. Comparisons between chronic CD clinical forms were performed using Mann-Whitney test and correlation Spearman's test. FH levels were correlated positively with triglycerides (P = .001, r = .39), LDL-C (P = .009, r = .3), cholesterol (P = .02, r = .28), uCRP (P = .029, r = .31) and BMI (P = .008, r = .34); and negatively with HDL-C (P = .03, r = -.25) levels. Dyslipidemic patients showed higher FH levels compared to normolipidemic, although no difference for FH levels was observed between chronic CD clinical forms. Alternative pathway of complement may be a link between immune response and metabolic disorders, with important immunoregulatory role in chronic CD.

Untargeted metabolomics for plasma biomarker discovery for early chronic kidney disease diagnosis in pediatric patients using LC-QTOF-MS.

Pediatric chronic kidney disease (CKD) is a clinical syndrome characterized by renal hypofunction occurring due to gradual and irreversible kidney damage that can further progress over time. New biomarkers may help early diagnosis of pediatric patients suffering from CKD and improve the outcome. Untargeted metabolomics based on LC-QTOF-MS has been used to find new biomarkers for the early diagnosis of CKD in plasma from pediatric patients. In order to avoid any bias in the determination of statistically significant entities as a consequence of the data analysis method followed, two different chemometric approaches have been used, Mass Profiler Professional (MPP) software and Matlab R2015a software. Metabolic fingerprints of control and CKD pediatric patients were compared and five metabolites which showed a significant change common to both data analysis procedures were identified. Sphingosine-1-phosphate, n-butyrylcarnitine, cis-4-decenoylcarnitine and an unidentified feature with 126.0930 m/z were found to be increased in plasma from pediatric patients with CKD, whereas bilirubin was significantly decreased. A partial least squares discriminant analysis model built with these 5 entities classified correctly 96% of the samples. In addition, when considering only early CKD patients against controls, a performance of 97% was obtained. Thus, these promising metabolites could be suitable biomarkers for the early diagnosis of pediatric CKD in a clinical setting.

Antimicrobial effectiveness of combinations of oxidant and chelating agents in infected dentine: an ex vivo confocal laser scanning microscopy study.

AIM: To evaluate the intratubular antimicrobial activity of several oxidant and chelating agents associated with or without surfactants in experimentally infected root canals, using confocal laser scanning microscopy. METHODOLOGY: Twenty-four dentine blocks from bovine incisors were contaminated for 5 days with Enterococcus faecalis (ATCC- 29212). Ten contaminated dentine specimens were irrigated for 5 min with 5.25% NaOCl followed by 17% EDTA for 2 min, and the other 10 with Hypoclean for 5 min followed by Tetraclean NA for 2 min. The remaining four specimens were used as positive and negative controls (two samples each). Then, dentine blocks were stained with Live/Dead BacLight for analysis of the remaining live or dead bacteria using confocal laser scanning microscopy. Comparison between and within groups was performed using the Mann-Whitney test for independent samples and the Wilcoxon signed-rank test, respectively. RESULTS: After exposure to irrigants, the positive control group had a median of 67.41% of viable bacteria (95% CI: 48.15, 78.9), whilst the NaOCl+EDTA group and Hypoclean+Tetraclean NA group had 3.77% (1.28, 15.92) and 0.87% (-0.42, 4.30) of viable bacteria, respectively. These results were significantly different from each other, both overall and distinct by region (cervical and medium third), or depth (superficial and deep layer) (P < 0.01 in all cases). CONCLUSIONS: The use of adjunctive agents reducing the surface tension associated with oxidant and chelating agents improved the antimicrobial activity of irrigating solutions and intratubular decontamination against Enterococcus faecalis, possibly due to better removal of the smear layer and deeper penetration into dentinal tubules.

Antibacterial properties of silver nanoparticles as a root canal irrigant against Enterococcus faecalis biofilm and infected dentinal tubules.

AIM: To evaluate the antimicrobial action of an irrigant containing silver nanoparticles in an aqueous vehicle (AgNp), sodium hypochlorite and chlorhexidine against Enterococcus faecalis biofilm and infected dentinal tubules. METHODOLOGY: Bovine dentine blocks were used for E. faecalis biofilm development for 21 days and irrigated with 94 ppm AgNp solution, 2.5% NaOCl and 2% chlorhexidine for 5, 15 and 30 min. For infection of dentinal tubules with E. faecalis, dentine specimens from bovine incisors were submitted to a contamination protocol over 5 days, with eight centrifugation cycles on every alternate day, and irrigated with the same solutions and time intervals used for the biofilm. The specimens were stained with the Live/Dead technique and evaluated using a confocal laser scanning microscope (CLSM). The bioImage_L software was used for measurement of the total biovolume of biofilm in µm3 and percentage of viable bacteria (green cells) in biofilm and in dentinal tubules found after the irrigation. Statistical analyses were performed using Kruskal-Wallis and Dunn's tests for quantification of viable cells in biofilm, the Friedman test for comparisons of viable bacteria in dentinal tubules in different areas of the root canal and the Mann-Whitney U-test to compare the action of the irrigants between the two methods (P < 0.05). RESULTS: The AgNp solution eliminated fewer bacteria, but was able to dissolve more biofilm compared with chlorhexidine (P < 0.05). NaOCl had the greatest antimicrobial activity and biofilm dissolution capacity. AgNp solution had less antimicrobial action in infected dentinal tubules compared with NaOCl (P < 0.05). The AgNp solution after 5 min was more effective in eliminating planktonic bacteria in dentinal tubules than in biofilm, but at 30 min fewer viable bacteria were observed in the biofilm compared with intratubular dentine (P < 0.05). CONCLUSIONS: AgNp irrigant was not as effective against E. faecalis compared to solutions commonly used in root canal treatment. NaOCl is appropriate as an irrigant because it was effective in disrupting biofilm and in eliminating bacteria in biofilms and in dentinal tubules.

Neuro-ophthalmologic manifestations in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that can affect different ocular structures, such as cornea, conjunctiva, episclera, sclera, uveal tract, retina, optic nerve and vessels. Neuro-ophthalmologic manifestations in SLE include different degrees of involvement of retina, choroid and optic nerve. Retinal changes are one of the most common ocular involvements and are frequently used as clinical criteria for activity, even if isolated. Studies show that up to 29% of patients with active SLE manifest retinal disease. The exact prevalence of choroidal disease is unknown, but is thought to be less common than retinopathy, due to under-diagnosis. Optic nerve disease, represented by optic neuritis and anterior/posterior ischaemic optic neuropathy, affects approximately 1% of SLE patients. These ocular manifestations have been associated with neurologic flares, antiphospholipid antibodies, nephropathy, and increased mortality. The aim of this paper is to review the different aspects of neuro-ophthalmologic involvement in SLE. Since these manifestations are frequent and potentially severe, a multi-professional team approach is needed to investigate properly and provide early aggressive treatment in order to avoid visual sequelae.

How functional disability relates to dentition in community-dwelling older adults in Brazil.

OBJECTIVE: To assess the association between dentition status and functional disabilities among community-dwelling older individuals. SUBJECTS AND METHODS: A total of 5012 individuals aged 65-74 were interviewed in 2013 during a nationwide community-based survey. This cross-sectional study used a questionnaire including information on dentition, locomotion impairment, difficulties in basic and instrumental activities of daily life, socio-demographic characteristics, behaviour and general health status. Poisson regression models assessed prevalence ratios (PR) and 95% confidence intervals (95% CI). RESULTS: About 31.3% of older people in Brazil reported having ≥21 preserved teeth, the condition for a functional dentition without prosthetic appliances. This proportion was significantly lower among individuals with locomotion impairment and difficulties in basic and instrumental activities of daily life. After adjusting for relevant covariates, functional dentition remained statistically associated with severe (PR = 0.69, 95% CI = 0.15-0.63) and total (PR = 0.69, 95% CI = 0.26-0.44) locomotion impairments, and difficulties in basic activities of daily life (PR = 0.70, 95% CI = 0.53-0.93). CONCLUSION: Having a functional dentition was significantly less prevalent among those affected by functional disabilities, and this association is unlikely to be due to insufficient control for relevant covariates. The association between dentition and functional disability is bidirectional and involves a complex interplay of one upon the other, and of factors that influence both outcomes.

Lipid-based nanovesicles for nanomedicine.

Molecular self-assembly has enabled the fabrication of biologically inspired, advanced nanostructures as lipid-based nanovesicles (L-NVs). The oldest L-NVs, liposomes, have been widely proposed as potential candidates for drug delivery, diagnostic and/or theranostic applications and some liposome-based drug products have already stepped from the lab-bench to the market. This success is attributed to their ability to encapsulate both hydrophobic and/or hydrophilic molecules, efficiently carry and protect them within the body and finally deliver them at the target site. These positive features are also coupled with high biocompatibility. However, liposomes still present some unsolved drawbacks, such as poor colloidal stability, short shelf-life, restricted and expensive conditions of preparation because of the inherent nature of their fundamental constituents (phospholipids). The new tools available in the self-assembly of controlled molecules have significantly advanced the field of L-NV design and synthesis, and non-liposomal L-NVs have been recently developed; this new generation of nanovesicles can represent a paradigm shift in nanomedicine: they may complement liposomes, showing their advantages and overcoming most of their drawbacks. Clearly, being still young, their rocky way to the clinic first and then to the market has just started and it is still long, but they have all the potentialities to reach their objective target. The purpose of this review is to first present the large plethora of L-NVs available, focusing on this new generation of non-liposomal L-NVs and showing their similarities and differences with respect to their ancestors (liposomes). Since the overspread of a nanomaterial to the market is also strongly dependent on the availability of technological-scale preparation methods, we will also extensively review the current approaches exploited for L-NV production. The most cutting-edge approaches based on compressed fluid (CF) technologies will be highlighted here since they show the potential to represent a game-change in the production of L-NVs, favouring their step from the bench to the market. Finally, we will briefly discuss L-NV applications in nanomedicine, looking also for their future perspectives.

Fisetin as a promising antifungal agent against Cryptocococcus neoformans species complex.

AIMS: The aim of this study was to investigate the mechanisms of action of fisetin, a flavonol with antifungal activity previously evaluated against the Cryptococcus neoformans species complex. METHODS AND RESULTS: Ergosterol content and flow cytometry analysis were determined for the C. neoformans species complex in the presence of fisetin and ultrastructural analysis of morphology was performed on Cryptococcus gattii and C. neoformans. Decrease in the total cellular ergosterol content after exposure to fisetin ranged from 25·4% after exposure to 128 µg ml(-1) to 21·6% after exposure to 64 µg ml(-1) of fisetin compared with the control (without fisetin). The fisetin effects obtained with flow cytometry showed metabolic impairment, and alterations in its normal morphology caused by fisetin in C. neoformans cells were verified using scanning electron microscopy. CONCLUSIONS: Fisetin is a compound that acts in the biosynthesis of ergosterol. Flow cytometry showed that fisetin reduced viability of the metabolically active cells of C. gattii, while morphological changes explain the action of fisetin in inhibiting growth of these fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: This study supports the idea that fisetin may represent a good starting point for the development of future therapeutic substances for cryptococcosis.

Sandwich consumption in relation to daily dietary intake and diet quality among US adults, 2003-2012.

OBJECTIVES: About one half of the US adult population consumed one or more sandwiches on any given day. Sandwich consumption may profoundly influence people's calorie intake and diet quality. However, few studies have investigated the nutritional impact of sandwich consumption at the national level. We examined sandwich consumption in relation to daily dietary intake and diet quality among US adults. STUDY DESIGN: Retrospective data analysis. METHODS: Nationally representative sample consisting of 27,075 adults 18 years of age and above came from the National Health and Nutrition Examination Survey (NHANES) 2003-2012 waves. Sandwich consumption was identified based on the What We Eat in America Food Category and the NHANES diet combination code. First-difference estimator addressed confounding bias from time-invariant unobservables (e.g. eating habits, taste preferences) by using within-individual variations in sandwich consumption between two non-consecutive 24-h dietary recalls. RESULTS: During 2003-2012, approximately 53.2% of US adults consumed sandwiches on any given day. Among sandwich consumers, nearly a quarter of daily total calorie intake and about a third of total fat, saturated fat, cholesterol, and sodium intake came from sandwich consumption. Men, non-Hispanic whites, young and middle-aged adults, people with higher income, and overweight/obese adults were more likely to consume sandwiches. Among those who consumed sandwiches on either but not both of the two dietary recall days, in comparison to no sandwich consumption, sandwich consumption on a dietary recall day was associated with an increase in daily intake of total energy by 98.7 kcal, total fat 6.8 g, and sodium 267.9 mg (P-value < 0.001). Statistically significant but relatively small differences were also found for other nutrients and food groups-sandwich consumption was associated with an increase in daily intake of sugar by 3.0 g (P-value < 0.01) and saturated fat 2.5 g (P-value < 0.001), a decrease in daily intake of vegetables by 8.4 g (P-value < 0.01), fruit 4.5 g (P-value < 0.05), and fibre 0.8 g (P-value < 0.001), and a decrease in the Healthy Eating Index-2010 score by 1.6 points (P-value < 0.001). CONCLUSIONS: Consumers should prudently evaluate the calorie/nutrient content of sandwich in order to make healthier dietary choices.