Lack of Association between Inadequate Micronutrient Intake and Prognosis in Outpatients with Heart Failure.

Inadequate nutrient intake can lead to worse outcomes in patients with heart failure (HF). This prospective cohort study aimed to assess the prevalence of inadequate micronutrient intake and their association with prognosis in 121 adult and elderly outpatients with HF. Habitual micronutrient intake was evaluated using 24-h dietary recalls (minimum 2 and maximum 6). Participants were grouped into moderate (n = 67) and high (n = 54) micronutrient deficiency groups, according to the individual assessment of each micronutrient intake. Patients' sociodemographic, clinical, and anthropometric data and clinical outcomes (hospitalization and mortality) within 24 months were collected. Overall and event-free survival rates were calculated using Kaplan-Meier estimates, and curves were compared using the log-rank test. The death risk rate (hazard ratio (HR)) was calculated using Cox's univariate model. The rate of inadequate intake was 100% for vitamins B1 and D and above 80% for vitamins B2, B9, and E, calcium, magnesium, and copper. No differences in overall survival and event-free survival were observed between groups of HF outpatients with moderate and high micronutrient deficiencies (HR = 0.94 (CI = 0.36-2.48), p = 0.91, and HR = 1.63 (CI = 0.68-3.92), p = 0.26, respectively), as well as when the inadequacy of each micronutrient intake was evaluated alone (all p > 0.05). In conclusion, a high prevalence of inadequate micronutrient intake was observed in outpatients with HF. Inadequate micronutrient intake was not associated with hospitalization and mortality in this group of patients.

The importance of combining methods to assess Candida albicans biofilms following photodynamic inactivation.

BACKGROUND: Methylene blue (MB)-mediated photodynamic inactivation (PDI) has shown good results in killing Candida spp. Although MB solutions are commonly used, new formulations have been designed to improve PDI. However, chemical substances in the formulation may interfere with the PDI outcome. In this sense, different methodologies should be used to evaluate PDI in vitro. Herein, we report different methodologies to evaluate the effects of PDI with an oral formulation (OF) containing 0.005% MB on Candida albicans biofilm. METHODS: Biofilms were treated using the MB-OF, with 5 min pre-irradiation time and exposure to a 640 nm LED device (4.7 J/cm2). PDI was evaluated by the XTT reduction test, counting the colony forming units (CFU), a filamentation assay, crystal violet (CV) staining, and scanning electronic microscopy (SEM). RESULTS: PDI was able to reduce around 1.5 log10 CFU/mL, even though no significant differences were noted in metabolic activity in comparison to the control immediately after PDI. A significant decrease in yeast to hyphae transition was observed after PDI, while the biofilm exhibited flattened cells and a reduced number of yeasts in SEM. The CV assay showed increased biomass. CONCLUSION: MB-OF-mediated PDI was effective in C. albicans biofilms, as it significantly reduced the CFU/mL and the virulence of surviving cells. The CV data were inconclusive, since the OF components interacted with the CV, making the data useless. Taken together, our data suggest that the association of different methods allows complementary responses to assess how PDI mediated by a formulation impacts biofilms.

Biomarkers of Zinc and Copper Status and Associated Factors in Outpatients with Ischemic and Non-Ischemic Heart Failure.

Background: Few studies have explored the impact of ischemic and non-ischemic etiologies of heart failure and other factors associated with heart failure on zinc and copper status. This study examined zinc and copper status in 80 outpatients with ischemic (n = 36) and non-ischemic (n = 44) heart failure and associations with biodemographic, clinical, biochemical, and nutritional parameters.Materials: Biomarkers of plasma zinc and copper, copper-zinc ratio, 24-h urinary zinc excretion, ceruloplasmin, and dietary intake of zinc and copper were assessed. Plasma zinc and copper and urinary zinc were measured by inductively coupled plasma mass spectrometry (ICP-MS).Results: Patients with ischemic heart failure showed lower dietary zinc intake and higher dietary copper intake (both p = 0.02). Zinc and copper in plasma, copper-zinc ratio, ceruloplasmin, and 24-h urinary zinc excretion showed no statistical differences between the groups (all p ≥ 0.05). An inverse association was found between age (ß =-0.001; p = 0.005) and the use of diuretics (ß = -0.047; p = 0.013) and plasma zinc. Copper levels in plasma (ß = 0.001; p < 0.001), and albumin (ß = 0.090; p<0.001) were directly associated with plasma zinc. A positive association was found between ceruloplasmin (ß = 0.011; p < 0.001), gamma-glutamyl transferase (ß = 0.001; p < 0.001), albumin (ß = 0.077; p = 0.001), and high-sensitivity c-reactive protein (ß = 0.001; p = 0.024) and plasma copper.Conclusion: Zinc and copper biomarkers in clinically stable patients with heart failure did not seem to be responsive to the differences in zinc and copper intake observed in this study, regardless of heart failure etiology. The predictors of plasma zinc and copper levels related to oxidative stress and inflammation should be monitored in heart failure clinical practice.

Vitamin D status and predictors of 25-hydroxyvitamin D levels in patients with heart failure living in a sunny region / Estado de la vitamina D y predictores de los niveles de 25-hidroxivitamina D en pacientes con insuficiencia cardíaca residentes en una región soleada

AIMS: hypovitaminosis D has frequently been identified in patients with heart failure (HF). However, few studies have been conducted in regions with high solar incidence. Therefore, this study aimed to evaluate vitamin D status and predictors of 25-hydroxyvitamin D (25(OH)D) levels in patients with HF living in a sunny region (5 °- 6 °S). METHODS: this cross-sectional study enrolled 70 patients with HF. Biodemographic, clinical, biochemical, dietary, and sun exposure data were collected, and 25(OH)D levels were measured. RESULTS: the mean 25(OH)D level was 40.1 (12.4) ng/mL, and 24.3 % (95 % CI: 14.2-33.8) of patients with HF had hypovitaminosis D (25(OH) D < 30 ng/mL). Female patients (p = 0.001), those with ischemic etiology (p = 0.03) and those with high parathyroid hormone levels (> 67 pg/mL) (p = 0.034) were more likely to present hypovitaminosis D. Higher 25(OH)D levels were observed in men than in women (β = 7.78, p = 0.005) and in patients with HF in New York Heart Association (NHYA) functional class I when compared to those in class III/IV (β = 8.23, p = 0.032). CONCLUSIONS: the majority of patients with HF had sufficient 25(OH)D levels. Sex and functional classification were identified as independent predictors of 25(OH)D levels. These results highlight the need for increased monitoring of vitamin D status among female patients with heart failure and those with more severe symptoms

OBJETIVOS: la hipovitaminosis D se ha identificado con frecuencia en pacientes con insuficiencia cardíaca (IC). Sin embargo, pocos estudios se han realizado en regiones con una alta exposición solar. Por lo tanto, este estudio tuvo como objetivo evaluar el estado de la vitamina D y los predictores de los niveles de 25-hidroxivitamina D (25(OH)D) en pacientes con IC que viven en una región soleada (5 °-6 °S). MÉTODOS: este estudio transversal incluyó a 70 pacientes con IC. Se recopilaron datos biodemográficos, clínicos, bioquímicos, dietéticos y de exposición solar, y se midieron los niveles de 25(OH)D. RESULTADOS: el nivel medio de 25(OH)D fue de 40,1 (12,4) ng/mL y el 24,3 % (IC 95 %: 14,2-33,8) de los pacientes con IC tenían hipovitaminosis D (25(OH)D < 30 ng/mL. Las pacientes mujeres (p = 0,001), aquellos con IC de etiología isquémica (p = 0,03) y aquellos otros pacientes con niveles altos de hormona paratiroidea (> 67 pg/mL) (p = 0,034) tenían más probabilidades de presentar hipovitaminosis D. Se observaron niveles más altos de 25(OH)D en los hombres que en las mujeres (β = 7,78, p = 0,005), y en los pacientes con IC de clase funcional I de la New York Heart Association (NHYA) que en los de clase III/IV (β = 8,23, p = 0,032). CONCLUSIONES: la mayoría de los pacientes con IC tenían niveles suficientes de 25(OH)D. El sexo y la clasificación funcional se identificaron como predictores independientes de los niveles de 25(OH)D. Estos resultados destacan la necesidad de un mayor control del estado de la vitamina D entre las mujeres con insuficiencia cardíaca y los pacientes con síntomas más graves

Vitamin D status and predictors of 25-hydroxyvitamin D levels in patients with heart failure living in a sunny region.

INTRODUCTION: Aims: hypovitaminosis D has frequently been identified in patients with heart failure (HF). However, few studies have been conducted in regions with high solar incidence. Therefore, this study aimed to evaluate vitamin D status and predictors of 25-hydroxyvitamin D (25(OH)D) levels in patients with HF living in a sunny region (5 °- 6 °S). Methods: this cross-sectional study enrolled 70 patients with HF. Biodemographic, clinical, biochemical, dietary, and sun exposure data were collected, and 25(OH)D levels were measured. Results: the mean 25(OH)D level was 40.1 (12.4) ng/mL, and 24.3 % (95 % CI: 14.2-33.8) of patients with HF had hypovitaminosis D (25(OH)D < 30 ng/mL). Female patients (p = 0.001), those with ischemic etiology (p = 0.03) and those with high parathyroid hormone levels (> 67 pg/mL) (p = 0.034) were more likely to present hypovitaminosis D. Higher 25(OH)D levels were observed in men than in women (ß = 7.78, p = 0.005) and in patients with HF in New York Heart Association (NHYA) functional class I when compared to those in class III/IV (ß = 8.23, p = 0.032). Conclusions: the majority of patients with HF had sufficient 25(OH)D levels. Sex and functional classification were identified as independent predictors of 25(OH)D levels. These results highlight the need for increased monitoring of vitamin D status among female patients with heart failure and those with more severe symptoms.

INTRODUCCIÓN: Objetivos: la hipovitaminosis D se ha identificado con frecuencia en pacientes con insuficiencia cardíaca (IC). Sin embargo, pocos estudios se han realizado en regiones con una alta exposición solar. Por lo tanto, este estudio tuvo como objetivo evaluar el estado de la vitamina D y los predictores de los niveles de 25-hidroxivitamina D (25(OH)D) en pacientes con IC que viven en una región soleada (5 °-6 °S). Métodos: este estudio transversal incluyó a 70 pacientes con IC. Se recopilaron datos biodemográficos, clínicos, bioquímicos, dietéticos y de exposición solar, y se midieron los niveles de 25(OH)D. Resultados: el nivel medio de 25(OH)D fue de 40,1 (12,4) ng/mL y el 24,3 % (IC 95 %: 14,2-33,8) de los pacientes con IC tenían hipovitaminosis D (25(OH)D < 30 ng/mL. Las pacientes mujeres (p = 0,001), aquellos con IC de etiología isquémica (p = 0,03) y aquellos otros pacientes con niveles altos de hormona paratiroidea (> 67 pg/mL) (p = 0,034) tenían más probabilidades de presentar hipovitaminosis D. Se observaron niveles más altos de 25(OH)D en los hombres que en las mujeres (ß = 7,78, p = 0,005), y en los pacientes con IC de clase funcional I de la New York Heart Association (NHYA) que en los de clase III/IV (ß = 8,23, p = 0,032). Conclusiones: la mayoría de los pacientes con IC tenían niveles suficientes de 25(OH)D. El sexo y la clasificación funcional se identificaron como predictores independientes de los niveles de 25(OH)D. Estos resultados destacan la necesidad de un mayor control del estado de la vitamina D entre las mujeres con insuficiencia cardíaca y los pacientes con síntomas más graves.

Factors associated with suicide risk among nurses and physicians: a cross-section study.

OBJECTIVE: to estimate the prevalence and factors associated with suicide risk among nurses and physicians. METHOD: a cross-sectional study carried out at a university hospital with 216 health professionals, who answered a socio-demographic-labor questionnaire, the Mini International Neuropsychiatric Interview (MINI) for assessing suicide risk, and the Depression, Anxiety and Stress Scale (DASS 21). The Poisson Regression Model was used for multiple analysis. RESULTS: it was identified that variables such as not having a partner, history of attempted suicide, stress and depression symptoms were statistically associated with suicide risk. The prevalence of lifelong suicide attempts among nurses was 9.41%, and among physicians, 2.29%. CONCLUSION: the findings of this investigation enable the understanding of suicidal behavior among hospital nurses and physicians, in addition to enabling the development of prevention strategies in order to reduce suicide risk prevalence in this population group.

Photodynamic antimicrobial chemotherapy action of phenothiazinium dyes in planktonic Candida albicans is increased in sodium dodecyl sulfate.

BACKGROUND: Photodynamic antimicrobial chemotherapy (PACT) is an important therapeutic platform for antimicrobial activity, especially due to the resistant strains, however, the physical-chemical properties of the photosensitizers may affect the final outcome. Recently, this research group showed that the control of aggregation potentiates the PACT action of Methylene Blue. In this study, the researchers investigated the PACT action of other phenothiazinium dyes (PDs), such as Azure A, Azure B (AB), and 1,9-dimethyl methylene blue (DMMB), and the effects of different mediums which modulate PD aggregation were studied. METHODS: The C. albicans planktonic culture was treated with PDs at different concentrations (0-100.00 mg/L), both in the dark (5 min) and irradiated (640 ±â€¯12 nm LED during 30 min, 2.60 mW/cm2). After the treatments, the diluted samples were grown on Sabouraud Dextrose Agar and maintained at 37 °C for 24 h. Later, one single concentration was defined and the inoculum was treated with the compounds in different media (water, phosphate buffer saline - PBS, physiological solution - 0.90 % NaCl, urea 1.00 mol/L, and sodium dodecyl sulfate 0.25 % - SDS), at 20.00 mg/L for AA and AB, and at 0.50 mg/L for DMMB. The cell uptake and the dimer to the monomer ratios were determined by spectrophotometry. RESULTS: PACT with the PDs showed a reduction in the CFU/mL, with DMMB being the most effective due to the higher cell uptake within the series. When PACT was applied in the different mediums, the inactivation response was influenced by the medium. In water, the physiological solution, PBS, and urea showed no significant differences in relation to the control group. On the other hand, SDS potentiated the PACT action, and the inactivation of the microorganism was achieved with AB and DMMB. CONCLUSIONS: In water, DMMB presented the highest PACT action within the PDs being studied, reaching a microorganism inactivation with a 1.00 mg/L solution, mainly due to its high cell uptake. SDS 0.25 % increased the PACT action of the PDs studied, mainly with DMMB and AB, and this may be used in formulations to develop antimicrobial treatments to be used in clinics.

Factors associated with suicide risk among nurses and physicians: a cross-section study / Factores asociados al riesgo de suicidio entre enfermeros y médicos: un estudio transversal / Fatores associados ao risco de suicídio entre enfermeiros e médicos: estudo transversal

ABSTRACT Objective: to estimate the prevalence and factors associated with suicide risk among nurses and physicians. Method: a cross-sectional study carried out at a university hospital with 216 health professionals, who answered a socio-demographic-labor questionnaire, the Mini International Neuropsychiatric Interview (MINI) for assessing suicide risk, and the Depression, Anxiety and Stress Scale (DASS 21). The Poisson Regression Model was used for multiple analysis. Results: it was identified that variables such as not having a partner, history of attempted suicide, stress and depression symptoms were statistically associated with suicide risk. The prevalence of lifelong suicide attempts among nurses was 9.41%, and among physicians, 2.29%. Conclusion: the findings of this investigation enable the understanding of suicidal behavior among hospital nurses and physicians, in addition to enabling the development of prevention strategies in order to reduce suicide risk prevalence in this population group.

RESUMEN Objetivo: estimar la prevalencia y los factores asociados al riesgo de suicidio entre enfermeros y médicos. Método: estudio transversal, desarrollado en un hospital universitario con 216 profesionales de la salud, quienes respondieron un cuestionario sociodemográfico-laboral; el Mini International Neuropsychiatric Interview (MINI), para evaluar el riesgo de suicidio; Escala de Depresión, Ansiedad y Estrés (DASS 21). El modelo de regresión de Poisson se utilizó para el análisis múltiple. Resultados: se identificó que variables como no tener pareja, antecedentes de intento de suicidio, síntomas de estrés y depresión se asociaron estadísticamente con el riesgo de suicidio. La prevalencia de intentos de suicidio de por vida entre enfermeras y médicos fue del 9,41% y del 2,29%. Conclusión: los hallazgos de esta investigación permiten comprender la conducta suicida entre enfermeros y médicos del área hospitalaria, además de posibilitar el desarrollo de estrategias de prevención con el fin de reducir la prevalencia de riesgo suicida en este grupo poblacional.

RESUMO Objetivo: estimar a prevalência e os fatores associados ao risco de suicídio entre enfermeiros e médicos. Método: estudo transversal, desenvolvido em um hospital universitário com 216 profissionais da saúde, que responderam a um questionário sociodemográfico-laboral; ao Mini International Neuropsychiatric Interview (MINI), para avaliação do risco de suicídio; Escala de Depressão, Ansiedade e Estresse (DASS 21). Utilizou-se para análise múltipla, o Modelo de Regressão de Poisson. Resultados: identificou-se que variáveis, como não ter companheiro, histórico de tentativa de suicídio, sintomas de estresse e depressão se associaram estatisticamente ao risco de suicídio. A prevalência das tentativas de suicídios ao longo da vida entre enfermeiros foi de 9,41% e médicos de 2,29%. Conclusão: Os achados desta investigação possibilitam a compreensão do comportamento suicida entre os enfermeiros e médicos da área hospitalar, além de viabilizar a elaboração de estratégias de prevenção com intuito de diminuir a prevalência do risco de suicídio nesse grupo populacional.

Guia prático para o manejo da dor / Guia prático para o manejo da dor

O estudo da dor e suas particularidades é de grande importância para o tratamento de diversas patologias e para a melhora na qualidade de vida dos pacientes. A maioria das disfunções orgânicas tem a dor como um ponto importante da sua manifestação. Dessa maneira, é justificável a elaboração de conteúdo para auxiliar os profissionais da saúde no entendimento e tratamento das principais causas de dores agudas e crônicas. Este livro foi elaborado com o objetivo de servir como um guia prático para o manejo da dor por profissionais e acadêmicos de Medicina. Engloba temas como conceitos e aspectos biopsicossociais da dor, além de questões mais complexas como a fisiologia da dor e o tratamento medicamentoso com o arsenal terapêutico existente. Finalmente, também trata dos diversos tipos de dor mais prevalentes e o conhecimento básico que envolve seu manejo.

Controlling methylene blue aggregation: a more efficient alternative to treat Candida albicans infections using photodynamic therapy.

Methylene Blue (MB) has been widely used in antimicrobial Photodynamic Therapy (aPDT), however, the mechanisms of action (Type I or Type II) are defined by its state of aggregation. In this sense, the identification of the relationships between aggregation, the mechanisms of action and the effectiveness against microorganisms, as well as the establishment of the means and the formulations that may favor the most effective mechanisms, are essential. Thus, the objective of this study was to assess the in vitro aPDT efficacies against Candida albicans, by using MB in vehicles which may influence the aggregation and present an oral formulation (OF) containing MB, to be used in clinical aPDT procedures. The efficacy of MB at 20 mg L-1 was tested in a range of vehicles (water, physiological solution - NaCl 0.9%, phosphate saline buffer - PBS, sodium dodecyl sulfate 0.25% - SDS and urea 1 mol L-1) in a C. albicans planktonic culture, when using 4.68 J cm-2 of 640 ± 12 nm LED for the irradiations, as well as 5 minutes of pre-irradiation time, together with measuring the UFC mL-1. Based upon these analyses, an OF containing MB in the most effective vehicle was tested in the biofilms, as a proposal for clinical applications. When comparing some of the vehicles, sodium dodecyl sulfate was the only one that enhanced an MB aPDT efficacy in a planktonic C. albicans culture. This OF was tested in the biofilms and 50 mg L-1 MB was necessary, in order to achieve some reduction in the cell viabilities after the various treatments. The light dosimetries still need further adaptations, in order for this formulation to be used in clinical applications. The present research has indicated that the development of this formulation for the control of MB aggregations may result in more effective clinical protocols.

Candida tropicalis affects the virulence profile of Candida albicans: an in vitro and in vivo study.

Candida albicans and Candida tropicalis are commensal microorganisms occurring in the oral cavity of approximately 50%-70% of healthy individuals. However, these microbes can become pathogenic through changes in the environment or weakened host immune system. Thus, the aim of this investigation was to evaluate the interaction between species of the genus Candida in the biofilm formation, filamentation, gene expression and virulence in Galleria mellonella. Coincubation of C. albicans with C. tropicalis cells after 48 h resulted in significant reduction of biofilm formation by decreasing viable cell counts, metabolic activity and hyphal growth. The C. albicans genes (BCR1, CPH1, EFG1, UME6, HWP1, ALS3, SAP5 and PLB2) were quantified by quantitative real-time polymerase chain reaction and most of genes were downregulated. Regarding in vivo assay, the groups that the larvae received C. albicans and C. tropicalis had a significant survival increase compared to the control group of C. albicans (P = 0.0001) in agreement with the in vitro results. In conclusion, C. tropicalis colonization was associated with a decrease in the growth of C. albicans, suggesting an antagonistic relation between these two species. Therefore, C. tropicalis by reducing C. albicans virulence profile may limit the ability of this pathogenic fungus to cause infection.

Photodynamic inactivation in the expression of the Candida albicans genes ALS3, HWP1, BCR1, TEC1, CPH1, and EFG1 in biofilms.

The objective of this study was to evaluate the effects of photodynamic inactivation (PDI) on Candida albicans biofilms, evaluating its effects on gene expression of ALS3, HWP1, BCR1, TEC1, CPH1, and EFG1 by yeast. Three samples of C. albicans were used in this study: a clinical sample from a patient with HIV (39S), a clinical sample from a patient with denture stomatitis lesion (Ca30), and a standard strain ATCC 18804. The quantification of gene expression was related to the production of those genes in the samples referred above using quantitative polymerase chain reaction (qPCR) assay in real time. The photosensitizer methylene blue at 300 uM and erythrosine at 400 uM, sensitized with low-power laser (visible red, 660 nm) and green LED (532 nm), respectively, were used for PDI. Four groups of each sample and PDI protocol were evaluated: (a) P+L+: sensitization with the photosensitizer and irradiation with light, (b) P+L-: only treatment with the photosensitizer, (c) P-L+: only irradiation with light, and (d) P-L-: without sensitization with the dye and absence of light. The results were analyzed by t test, with a significance level of 5%. The photodynamic inactivation was able to reduce the expression of all genes for both treatments, laser and LED. The fold-decrease for the genes ALS3, HWP1, BCR1, TEC1, CPH1, and EFG1 were 0.73, 0.39, 0.77, 0.71, 0.67, and 0.60 for laser, respectively, and 0.66, 0.61, .050, 0.43, 0.54, and 0.66 for LED, respectively. It could be concluded that PDI showed a reduction in the expression of C. albicans genes, suggesting its virulence decrease.

Study of Microbial Interaction Formed by "Candida krusei" and "Candida glabrata": "In Vitro" and "In Vivo" Studies.

Recently, the non-albicans Candida species have become recognized as an important source of infection and oral colonization by association of different species in a large number of immunosuppressed patients. The objective of this study was to evaluate the interactions between C. krusei and C. glabrata in biofilms formed in vitro and their ability to colonize the oral cavity of mouse model. Monospecies and mixed biofilms were developed of each strain, on 96-well microtiter plates for 48 h. These biofilms were analyzed by counting colony-forming units (CFU/mL) and by determining cell viability, using the XTT hydroxide colorimetric assay. For the in vivo study, twenty-four mice received topical applications of monospecie or mixed suspensions of each strain. After 48 h, yeasts were recovered from the mice and quantified by CFU/mL count. In the biofilm assays, the results for the CFU/mL count and the XTT assay showed that the two species studied were capable of forming high levels of in vitro monospecie biofilm. In mixed biofilm, the CFU of C. krusei increased (p=0.0001) and C. glabrata decreased (p=0.0001). The metabolic activity observed in XTT assay of mixed biofilm was significantly reduced compared with a single C. glabrata biofilm (p=0.0001). Agreeing with CFU in vitro count, C. glabrata CFU/mL values recovered from oral cavity of mice were statistically higher in the group with single infection (p=0.0001) than the group with mixed infection. We concluded that C. krusei inhibits C. glabrata and takes advantage to colonize the oral cavity and to form biofilms.

Study of Microbial Interaction Formed by "Candida krusei" and "Candida glabrata": "In Vitro" and "In Vivo" Studies

Abstract Recently, the non-albicans Candida species have become recognized as an important source of infection and oral colonization by association of different species in a large number of immunosuppressed patients. The objective of this study was to evaluate the interactions between C. krusei and C. glabrata in biofilms formed in vitro and their ability to colonize the oral cavity of mouse model. Monospecies and mixed biofilms were developed of each strain, on 96-well microtiter plates for 48 h. These biofilms were analyzed by counting colony-forming units (CFU/mL) and by determining cell viability, using the XTT hydroxide colorimetric assay. For the in vivo study, twenty-four mice received topical applications of monospecie or mixed suspensions of each strain. After 48 h, yeasts were recovered from the mice and quantified by CFU/mL count. In the biofilm assays, the results for the CFU/mL count and the XTT assay showed that the two species studied were capable of forming high levels of in vitro monospecie biofilm. In mixed biofilm, the CFU of C. krusei increased (p=0.0001) and C. glabrata decreased (p=0.0001). The metabolic activity observed in XTT assay of mixed biofilm was significantly reduced compared with a single C. glabrata biofilm (p=0.0001). Agreeing with CFU in vitro count, C. glabrata CFU/mL values recovered from oral cavity of mice were statistically higher in the group with single infection (p=0.0001) than the group with mixed infection. We concluded that C. krusei inhibits C. glabrata and takes advantage to colonize the oral cavity and to form biofilms.

Resumo Recentemente, as espécies não albicans tem se tornado uma importante fonte de infecção e de colonização oral pela associação de espécies em um grande número de pacientes imunossuprimidos. O objetivo desse estudo foi avaliar a interação entre C. krusei e C. glabrata em biofilmes formados in vitro e sua capacidade em colonizar a cavidade oral em modelo de camundongo. Biofilmes monoespécies e mistos foram formados em placas de 96 poços por 48 h. Esses biofilmes foram analisados pela contagem de UFC/mL e pela determinação da viabilidade celular, usando ensaio de XTT. Para o estudo in vivo, vinte e quatro camundongos receberam aplicações tópicas de suspensões monoespécies e mistas de cada espécie. Após 48 h, as leveduras foram recuperadas dos camundongos e quantificadas por UFC/mL. Nos ensaios de biofilme, os resultados da contagem de UFC/mL e do ensaio de XTT mostraram que as duas espécies estudadas foram capazes de formar grande quantidade de biofilme monoespécie in vitro. Nos biofilmes mistos, a UFC/mL de C. krusei aumentou (p=0,0001) e de C. glabrata diminuiu (p=0,0001). A atividade metabólica observada no ensaio de XTT nos biofilmes mistos foi significantemente reduzida comparada com o biofilme formado apenas de C. glabrata (p=0,0001). Concordado com as contagens in vitro, os valores de UFC/mL de C. glabrata recuperados da cavidade oral dos camundongos foram estatisticamente maior no grupo com infecção simples (p=0,0001) do que do grupo com infecção mista. Nós concluímos que C. krusei inibe C. glabrata e possui vantagem em colonizar a cavidade oral e formar biofilmes.

Impact of multi-functional fermented goat milk beverage on gut microbiota in a dynamic colon model.

The aim of this research was to evaluate the effect of grape probiotic fermented beverages made of goat milk, with or without added grape pomace on gut microbiota in a Simulator of Human Intestinal Microbial Ecosystem (SHIME®). SHIME® model was used to investigate to assess changes in microbial composition and fermentation metabolites (short- and branched-chain fatty acids and ammonium), as well as under the antioxidant capacity. The results demonstrated that the beverages formulated, with or without grape pomace extract, exhibited high dietary fiber, oleic acid, phenolic compounds content and antioxidant activity. Both beverages also kept L. rhamnosus and S. thermophilus viable during their passage through the intestinal tract and had a positive effect on gut microbiota metabolism, increasing the antioxidant capacity and the production of short-chain fatty acids, and decreasing the ammonium concentration. Therefore, the multifunctional beverages formulated in this study can offer a new perspective for the production of foods with positive potential effects on human health.

Influence of sucrose on growth and sensitivity of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans to photodynamic therapy.

This study has evaluated the effects of photodynamic inactivation (PDI) using erythrosine as photosensitizer and green light-emitting diode (LED) on biofilms of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans. We have also evaluated the effect of sucrose on biofilm formation and bacterial growth and sensitivity to PDI. Biofilms were formed in suspension of 106 cells/ml on plates before being grown in broth culture with and without sucrose and incubated for 48 h. Next, the treatment was applied using erythrosine at a concentration of 400 µM for 5 min and green LED (532 ± 10 nm) for 3 min on biofilms alone and in combination. The plates were washed and sonicated to disperse the biofilms, and serial dilutions were carried and aliquots seeded in Sabouraud agar before incubation for 48 h. Next, the colony-forming units per milliliter (CFU/ml; log10) were counted and analyzed statistically (ANOVA, Tukey test, P ≤ 0.05). Results show that S. mutans favors the growth of C. albicans in biofilms with sucrose, with treatment not being effective. However, when the biofilm was grown without sucrose, we found a reduction in biofilm formation and a significant decrease in the PDI treatment (P < 0.0001). In conclusion, both growth and sensitivity to PDI in biofilms of C. albicans are strongly influenced by bacterial combination, and the presence of sucrose affected directly the growth and sensitivity of the biofilm to PDI as sucrose is the substrate for construction of the exopolysaccharide matrix.

Angiossarcoma primário de mama: desa o diagnóstico / Primary breast angiosarcoma: a diagnostic challenge

Os autores expõem um caso de uma mulher de 57 anos que apresentou uma massa palpável na mama direita. Na mamogra a, foi identi cada uma assimetria focal com gordura de permeio, sem achado suspeito à ultrassonogra a. A ressonância magnética das mamas, com meio de con­ traste, demonstrou um nódulo irregular e mal de nido, com baixo sinal em T1 sem supressão de gordura e alto sinal em T2 com supressão de gordura. Apresentou também realce heterogêneo irregular com septos espessos realçantes medindo 5,3 cm, cinética inicial rápida e curva do tipo persistente. Uma nova ultrassonogra a dirigida foi realizada, sendo observado um nódulo hi­ poecoico de 1,2 cm, associado à alteração da ecogenicidade do parênquima mamário adjacente. Realizou­se biópsia a vácuo do nódulo hipoecoico e do parênquima adjacente, guiada por ultras­ som. O estudo histopatológico demonstrou angiossarcoma primário da mama de baixo grau. O PET­CT, realizado 6 meses após a cirurgia, não evidenciou recidiva ou metástases.

The authors display a case of a woman aged 57 years who presented a palpable mass in the right breast. The mammography revelled a focal asymmetry, with no suspicious ndings on ultrasound. Breast MRI showed an irregular and unde ned mass with low signal intensity on T1 without fat suppression and high signal on T2­weighted fat­suppressed, with irregular heterogeneous enhancement with thick enhancement septa, fast initial kinetic enhancement and persistent type curve, measuring 5.3 cm. A new ultrasound exam was performed, observing hypoechoic nodule of 1.2 cm associated with changes in echogenicity of the adjacent breast parenchyma. Ultrasound­guided vacuum biopsy of the hypoechoic nodule and the adjacent parenchyma was performed. Histopathological study showed primary breast angiosarcoma of low­grade. The PET­CT performed six months after surgery showed no recurrence or metastasis.

Candida krusei and Candida glabrata reduce the filamentation of Candida albicans by downregulating expression of HWP1 gene.

Pathogenicity of Candida albicans is associated with its capacity switch from yeast-like to hyphal growth. The hyphal form is capable to penetrate the epithelial surfaces and to damage the host tissues. Therefore, many investigations have focused on mechanisms that control the morphological transitions of C. albicans. Recently, certain studies have showed that non-albicans Candida species can reduce the capacity of C. albicans to form biofilms and to develop candidiasis in animal models. Then, the objective of this study was to evaluate the effects of Candida krusei and Candida glabrata on the morphogenesis of C. albicans. Firstly, the capacity of reference and clinical strains of C. albicans in forming hyphae was tested in vitro. After that, the expression of HWP1 (hyphal wall protein 1) gene was determined by quantitative real-time PCR (polymerase chain reaction) assay. For both reference and clinical strains, a significant inhibition of the hyphae formation was observed when C. albicans was incubated in the presence of C. krusei or C. glabrata compared to the control group composed only by C. albicans. In addition, the culture mixed of C. albicans-C. krusei or C. albicans-C. glabrata reduced significantly the expression of HWP1 gene of C. albicans in relation to single cultures of this specie. In both filamentation and gene expression assays, C. krusei showed the higher inhibitory activity on the morphogenesis of C. albicans compared to C. glabrata. C. krusei and C. glabrata are capable to reduce the filamentation of C. albicans and consequently decrease the expression of the HWP1 gene.

Expressão dos genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 de Candida albicans em biofilmes após inativação fotodinâmica / Expression of the Candida albicans genes ALS3, HWP1, BCR1, TEC1, CPH1 and EFG1 in biofilms after photodynamic inactivation

Os micro-organismos estão se tornando cada vez mais resistentes aos antimicrobianos e cepas de Candida albicans resistentes aos antifúngicos tem sido isoladas, assim, torna-se importante e necessário a realização de pesquisas que avaliem os efeitos de novos métodos terapêuticos, como a inativação fotodinâmica antimicrobiana (aPDI). Assim, o objetivo deste estudo foi verificar os efeitos da inativação fotodinâmica sobre biofilmes de Candida albicans, avaliando seus efeitos sobre a expressão dos genes TEC1 (fator de transcrição), HWP1 (proteína de parede celular das hifas), EFG1 (regulador transcricional relacionado com a morfogênese), BCR1 (regulador da formação de biofilme e da parede celular), CPH1 (regulador transcricional envolvido na morfogênese) e ALS3 (adesina) de C. albicans. Foram avaliadas 30 amostras isoladas de pacientes portadores de HIV e 30 amostras de pacientes com estomatite protética, quanto a produção de biofilme, peso seco e filamentação. Destas, foram selecionadas as amostras mais virulentas de cada grupo que apresentaram melhor capacidade de formação de biofilme e filamentação. Assim, foi utilizada uma amostra clínica de C. albicans isolada de paciente portador de HIV, uma amostra clínica de C. albicans isolada de paciente com estomatite protética e uma cepa padrão ATCC 18804. A quantificação da expressão dos genes foi relacionada à produção desses genes nas amostras clínicas e na cepa de referência utilizando-se ensaio de PCR em tempo real. Para a aPDI, foram utilizados os fotossensibilizadores azul de metileno a 300 µM e eritrosina a 400 µM sensibilizados com laser de Índio-Gálio-Alumínio-Fósforo de baixa potência (vermelho visível, 660 nm) e LED verde (532 ± 10 nm), respectivamente. Foram avaliados quatro grupos experimentais para a aPDI: a) F+L+: sensibilização com o corante e irradiação com luz; b) F+L-: somente tratamento com o fotossensibilizador; c) F-L+: somente irradiação com luz e d) F-L-: sem sensibilização com o corante e ausência de luz. Os resultados foram analisados por t-test, com um nível de significância de 5%. Após a análise fenotípica, as amostras Ca30 e 39 S foram selecionadas para a realização da aPDI. Como esperado, apenas para o grupo F+L+, quando comparado com o grupo F-L-, todos os genes analisados foram sub expressos após a aPDI. O fold-decrease para os genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 foram 0,73; 0,39; 0,77; 0,71; 0,67 e 0,60; para laser, respectivamente, e 0,66; 0,61; 0,50; 0,43; 0,54 e 0,66; para LED, respectivamente. Pode-se concluir que a aPDI mostrou uma redução na expressão dos genes de C. albicans, sugerindo a diminuição de sua virulência(AU)

Os micro-organismos estão se tornando cada vez mais resistentes aos antimicrobianos e cepas de Candida albicans resistentes aos antifúngicos tem sido isoladas, assim, torna-se importante e necessário a realização de pesquisas que avaliem os efeitos de novos métodos terapêuticos, como a inativação fotodinâmica antimicrobiana (aPDI). Assim, o objetivo deste estudo foi verificar os efeitos da inativação fotodinâmica sobre biofilmes de Candida albicans, avaliando seus efeitos sobre a expressão dos genes TEC1 (fator de transcrição), HWP1 (proteína de parede celular das hifas), EFG1 (regulador transcricional relacionado com a morfogênese), BCR1 (regulador da formação de biofilme e da parede celular), CPH1 (regulador transcricional envolvido na morfogênese) e ALS3 (adesina) de C. albicans. Foram avaliadas 30 amostras isoladas de pacientes portadores de HIV e 30 amostras de pacientes com estomatite protética, quanto a produção de biofilme, peso seco e filamentação. Destas, foram selecionadas as amostras mais virulentas de cada grupo que apresentaram melhor capacidade de formação de biofilme e filamentação. Assim, foi utilizada uma amostra clínica de C. albicans isolada de paciente portador de HIV, uma amostra clínica de C. albicans isolada de paciente com estomatite protética e uma cepa padrão ATCC 18804. A quantificação da expressão dos genes foi relacionada à produção desses genes nas amostras clínicas e na cepa de referência utilizando-se ensaio de PCR em tempo real. Para a aPDI, foram utilizados os fotossensibilizadores azul de metileno a 300 µM e eritrosina a 400 µM sensibilizados com laser de Índio-Gálio-Alumínio-Fósforo de baixa potência (vermelho visível, 660 nm) e LED verde (532 ± 10 nm), respectivamente. Foram avaliados quatro grupos experimentais para a aPDI: a) F+L+: sensibilização com o corante e irradiação com luz; b) F+L-: somente tratamento com o fotossensibilizador; c) F-L+: somente irradiação com luz e d) F-L-: sem sensibilização com o corante e ausência de luz. Os resultados foram analisados por t-test, com um nível de significância de 5%. Após a análise fenotípica, as amostras Ca30 e 39 S foram selecionadas para a realização da aPDI. Como esperado, apenas para o grupo F+L+, quando comparado com o grupo F-L-, todos os genes analisados foram sub expressos após a aPDI. O fold-decrease para os genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 foram 0,73; 0,39; 0,77; 0,71; 0,67 e 0,60; para laser, respectivamente, e 0,66; 0,61; 0,50; 0,43; 0,54 e 0,66; para LED, respectivamente. Pode-se concluir que a aPDI mostrou uma redução na expressão dos genes de C. albicans, sugerindo a diminuição de sua virulência(AU)

Antibiofilm activity in vitro of Rosmarinus officinalis and Syzygium cumini glycolic extracts on Staphylococcus spp. of dentistry interest / Atividade antibiofilme in vitro dos extratos glicólicos de Rosmarinus officinalis e Syzygium cumini em Staphylococcus spp. de interesse odontológico

Objectives: The aim of this study was to identify the slime production and evaluate the effects of Rosmarinus officinalis (rosemary) and Syzygium cumini (jambolan) glycolic extracts, and 0.12% chlorhexidine (CHX) in biofilms formed by strains of coagulase-positive Staphylococcus - CPS and coagulase negative Staphylococcus - CNS isolated from the oral cavity. Material and Methods: Slime production was evaluated by two methods: the color of colony presented in Congo red agar, and through the amount of slime adhered to polystyrene. Biofilms were grown in acrylic resin discs immersed in broth, inoculated with microbial suspension (106 cells/ml) and incubated at 37°C/48 h. After formation, the biofilms were exposed for 5 minutes to glycol extracts, CHX or saline solution. The viability of biofilms was determined by counting the colony-forming units per milliliter (CFU/ml) in agar, and analyzed statistically by Tukey test (p< 0.05). Results: The strains S. aureus, S. schleiferi and S. epidermidis obtained the highest values of slime adhered to polystyrene. R. officinalis promoted reductions ranging from 12.1% to 78.7% in biofilms formed by isolates of CPS, and 9.2% to 73.7% in the biofilms of CNS. S. cumini reduced 12% to 55.7% in biofilms of CPS, and 7.9% to 71.5% in biofilms of CNS. With exception of S. saprophyticus, glycol extracts produced significant reductions in biofilms. For five isolates studied, R. officinalis produced greater reductions than CHX. Conclusion: R. officinalis and S. cumini showed effective antibiofilm activity against isolates that showed slime production.(AU)

Objetivos: O objetivo deste estudo foi identificar a produção de slime e avaliar os efeitos dos extratos glicólicos de Rosmarinus officinalis (alecrim), Syzygium cumini (jambolão) e 0,12% de clorexidina (CLX) em biofilmes formados por cepas de Staphylococcus coagulase positivo (SCP) e Staphylococcus coagulase negativo (SCN) da cavidade oral. Material e Métodos: A produção de slime foi avaliada por dois métodos: a cor da colônia apresentada em ágar vermelho Congo e pela quantidade de slime aderido ao poliestireno. Os biofilmes foram crescidos em discos de resina acrílica imersos em caldo, inoculados com suspensão microbiana (106 células/ml) e incubados a 37°C/48h. Após a formação, os biofilmes foram expostos durante 5 minutos aos extractos glicólicos, CLX ou solução salina. A viabilidade dos biofilmes foi determinada pela contagem das unidades formadoras de colônias por mililitro (UFC/ml) em ágar e analisada estatisticamente pelo teste de Tukey (p< 0,05). Resultados: As cepas S. aureus, S. schleiferi e S. epidermidis obtiveram os maiores valores de aderência ao poliestireno. R. officinalis promoveu reduções variando de 12,1% a 78,7% em biofilmes formados por isolados de SCP e 9,2% a 73,7% nos biofilmes de SCN. S. cumini reduziu de 12% a 55,7% nos biofilmes de SCP, e 7,9% a 71,5% nos biofilmes de SCN. Com exceção de S. saprophyticus, os extratos glicólicos produziram reduções estatísticas nos biofilmes. Para cinco isolados estudados, R. officinalis produziu maiores reduções do que CLX. Conclusão: R. officinalis e S. cumini mostraram atividade antibiofilme efetiva contra isolados que apresentaram produção de slime.(AU)