Fluconazole induces genotoxicity in cultured human peripheral blood mononuclear cells via immunomodulation of TNF-α, IL-6, and IL-10: new challenges for safe therapeutic regimens.

Context: Fluconazole (FNZ) is a drug used in antifungal therapy. However, the minimum FNZ dose to interfering with immune responses or inducing DNA damage is still unknown. Objective: This study investigated the toxicological profile of FNZ on cultured human peripheral blood mononuclear cells (PBMCs) treated with different concentrations of this azole. Materials and methods: Cultured PBMCs were exposed to FNZ (6, 12, 30, 60 and 120 µg/mL) and the toxicological profile was assessed by the following parameters: cytotoxic and nuclear division index (necrotic, apoptotic and viable cells), DNA damage (alkaline comet test), mutagenic potential (micronucleus test), cytokine modulation (IL-1, IL-6, IL-10, TNF-α, IFN-γ), and predictive toxicity (Osiris® and LAZAR® programs). Results: Our results demonstrated that FNZ induced cellular DNA damage and mutagenicity at concentrations above the plasma peak (>30 µg/mL) and 6 µg/mL, respectively, which was associated with increased TNF-α, and decrease IL-6 and IL-10 concentrations. These effects may be related to increased apoptosis and cytotoxic nuclear division index in the cultured PBMCs. In silico results indicated potential mutagenic, tumorigenic, irritant, and carcinogenic effects, which were partially confirmed by the above assays. Discussion and conclusions: Together, these findings suggest the need to rationalize the use of FNZ, especially if it is used for long periods or with concomitant pathologies requiring azole therapy that may increase FNZ's plasma concentration.

Imidazolium salts with antifungal potential against multidrug-resistant dermatophytes.

AIMS: To investigate the antidermatophytic action of a complementary set imidazolium salts (IMS), determining structure-activity relationships and characterizing the IMS toxicological profiles. METHODS AND RESULTS: The susceptibility evaluation of 45 dermatophytic clinical isolates, treated in vitro with eleven different IMS (ionic compounds) and commercial antifungals (nonionic compounds), was performed by broth microdilution, following the standard norm of CLSI M38-A2. All dermatophytes were inhibited by IMS, where the lowest minimum inhibitory concentration (MIC) values were observed for salts with n-hexadecyl segment in the cation side chain, containing either the chloride or methanesulfonate anion. 1-n-Hexadecyl-3-methylimidazolium chloride (C16 MImCl) and 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16 MImMeS) acted as fungicides, even in extremely low concentrations, wherein C16 MImMeS exerted this effect on 100% of the tested dermatophytes. Some of these IMS provoked evident alterations on the fungi cell morphology, causing a total cell damage of ≥ 70%. Importantly, none of the screened IMS were cytotoxic, mutagenic or genotoxic to human leucocyte cells. CONCLUSIONS: This report demonstrates for the first time the strong antifungal potential of IMS against multidrug-resistant dermatophytes, without presenting toxicity to human leucocyte cells at MIC. SIGNIFICANCE AND IMPACT OF THE STUDY: The expressive antifungal activity of IMS, combined with the in vitro nontoxicity, makes them promising compounds for the safe and effective treatment of dermatophytoses, mainly when this skin mycosis is unresponsive to conventional drugs.

Different risk factors for candidemia occur for Candida species belonging to the C. parapsilosis complex.

Previous studies have demonstrated reduced virulence in the species that comprise the Candida parapsilosis complex. We investigated a cohort of 93 patients with candidemia caused by this complex. Most infections were caused by C. parapsilosis (80.6%), followed by C. orthopsilosis (18.3%) and C. metapsilosis (1.1%). Renal failure (P < 0.001) and chronic liver diseases (P = 0.019) were more frequently encountered with infections caused by the C. orthopsilosis group, suggesting an association with patients who had a greater state of immune suppression in comparison with infections caused by C. parapsilosis sensu stricto.

Feeding preference of the shredder Phylloicus sp. for plant leaves of Chrysophyllum oliviforme or Miconia chartacea after conditioning in streams from different biomes.

Macroinvertebrate shredders consume preferably leaves conditioned by fungi and bacteria which offer greater palatability to them. Plant species in Cerrado present high concentration of chemical elements such as lignin and cellulose, phenols and tanins thus making them less attractive for shredders consumption and limiting the palatability. This study aimed to evaluate the feeding preference of a macroinvertebrate shredder of the genus Phylloicus for plant material from two different biomes (Cerrado and Mata Atlântica), after conditioning in a stream of Mata Atlântica and observing their physical and chemical characteristics. Senescent leaves were collected, monthly from the litterfall of riparian vegetation in a 500 m stretch of a stream in each biome from August 2014 to January 201. The most abundant species in each stream was selected for the experiment. The experimental design consisted in with two treatments. The first (T1) comprised leaf discs from Chrysophyllum oliviforme (Cerrado species) together with leaf discs of Miconia chartacea (Atlantic Forest species) which were conditioned in the Atlantic Forest stream. The second treatment (T2) involved leaf discs of Miconia chartacea conditioned in Mata Altlântica and Cerrado streams. Both tests had showed significant differences between the two treatments (T1 and T2). For T1, there was consumption of M. chartacea leaf discs by Phylloicus sp., but there was no consumption of C. oliviforme discs. For T2, there was preference for M. chartaceae leaves conditioned in a stream of Mata Atlântica than in Cerrado stream. The results showed that Phylloicus sp., had presented preference for food detritus of the Mata Antlântica biome and rejection to the one from Cerrado biome.

Anti-dermatophyte activity of Leguminosae plants from Southern Brazil with emphasis on Mimosa pigra (Leguminosae).

BACKGROUND: Intensive prophylactic use of antifungals leads to the increase of drug resistance and the need for new and more effective treatments are real. Plants from Leguminosae family are rich in flavonoids, for which numerous biological activities have been described, including antifungal effects. PURPOSE: To screen methanolic extracts from Leguminosae species looking for alternative sources for antifungal agents (anti-dermatophyte and anti-Candida) and their innocuity. METHODS: Antifungal activity was evaluated using the strains Candida albicans, C. krusei, C. glabrata, C. tropicalis, C. parapsilosis, Epidermophyton floccosum, Trichophyton mentagrophytes, T. rubrum and, Microsporum gypseum in the broth microdilution method. Later, the minimum inhibitory concentration (MIC) for Mimosa pigra, Eriosema heterophyllum, and Chamaecrista nictitans was determined. The most promising extract was fractionated and cytotoxicity and genotoxicity of the most active fraction were also assayed. RESULTS: Fungicide and/or fungistatic activity against dermatophyte strains were presented by 60% of the methanolic extracts assayed. M. pigra, E. heterophyllum, and C. nictitans methanolic extracts could inhibit dermatophyte strains at concentrations ranging from 1.9 to 1000µg/mL. M. pigra showed the lowest MIC values for a dichloromethane fraction (1.9µg/mL) without DNA damage at 10 and 50µg/mL and 100% of cell viability of human leukocytes. CONCLUSION: Our results indicate that methanolic extracts from Leguminosae plants are potential sources of antifungal compounds, mainly the extract and fractions from M. pigra. The dichloromethane fraction from M. pigra did not showed in vitro toxicity according to the applied assays.

Biodiesel production using fatty acids from food industry waste using corona discharge plasma technology.

This article aims to describe an alternative and innovative methodology to transform waste, frying oil in a potential energy source, the biodiesel. The biodiesel was produced from fatty acids, using a waste product of the food industry as the raw material. The methodology to be described is the corona discharge plasma technology, which offers advantages such as acceleration of the esterification reaction, easy separation of the biodiesel and the elimination of waste generation. The best conditions were found to be an oil/methanol molar ratio of 6:1, ambient temperature (25 °C) and reaction time of 110 min and 30 mL of sample. The acid value indicates the content of free fatty acids in the biodiesel and the value obtained in this study was 0.43 mg KOH/g. Peaks corresponding to octadecadienoic acid methyl ester, octadecanoic acid methyl ester and octadecenoic acid methyl ester, from the biodiesel composition, were identified using GC-MS. A major advantage of this process is that the methyl ester can be obtained in the absence of chemical catalysts and without the formation of the co-product (glycerin).

Analysis of cellular adhesion on superhydrophobic and superhydrophilic vertically aligned carbon nanotube scaffolds.

We analyzed GFP cells after 24h cultivated on superhydrophilic vertically aligned carbon nanotube scaffolds. We produced two different densities of VACNT scaffolds on Ti using Ni or Fe catalysts. A simple and fast oxygen plasma treatment promoted the superhydrophilicity of them. We used five different substrates, such as: as-grown VACNT produced using Ni as catalyst (Ni), as-grown VACNT produced using Fe as catalyst (Fe), VACNT-O produced using Ni as catalyst (NiO), VACNT-O produced using Fe as catalyst (FeO) and Ti (control). The 4',6-diamidino-2-phenylindole reagent nuclei stained the adherent cells cultivated on five different analyzed scaffolds. We used fluorescence microscopy for image collect, ImageJ® to count adhered cell and GraphPad Prism 5® for statistical analysis. We demonstrated in crescent order: Fe, Ni, NiO, FeO and Ti scaffolds that had an improved cellular adhesion. Oxygen treatment associated to high VACNT density (group FeO) presented significantly superior cell adhesion up to 24h. However, they do not show significant differences compared with Ti substrates (control). We demonstrated that all the analyzed substrates were nontoxic. Also, we proposed that the density and hydrophilicity influenced the cell adhesion behavior.

Extracellular matrix alterations during endochondral ossification in humans.

Immunohistochemical methods were employed to examine alterations in the cartilage extracellular matrix constituents associated with endochondral ossification in humans. The distributions of chondroitin 4- and 6-sulfate and keratan sulfate proteoglycan (PG) determinants, cartilage PG link protein, collagen types I and II, and fibronectin were determined in iliac crest growth-plate specimens using the avidin-biotin-horseradish peroxidase system. Collagen type II was distributed throughout the growth plate, providing a framework within which chondrocytes divided and formed clusters of differentiating (hypertrophic) cells. The septa between these clusters and their subchondral extensions into underlying bone trabeculae were rich in PG, PG link protein, and collagen type II and resembled the extracellular matrix of reserve cartilage. The territorial matrix associated with the differentiating cells within the clusters contained reduced amounts of collagen type II, PG link protein, and possibly cartilage PG. Collagen type I and fibronectin were detected within the cytoplasm of the maturing and degenerating cells, and fibronectin localized intensely to the pericellular matrix envelopes of these cells. These alterations presumably facilitate the degradation of the matrix associated with the cell clusters by invading vascular tissue, while the septa, which retain the characteristics of more typical cartilage matrix, are not degraded and firmly anchor the cartilage to the subchondral bone.

Natural and experimental infection of planorbids from the Island of Santa Catarina (Brazil).

Studies of eight localities on the Island of Santa Catarina revealed the presence of three species of the molluscan family Planorbidae: Biomphalaria tenagophila, Drepanotrema cimex and Biomphalaria oligoza, the first one being naturally infected by Cercaria ocellifera, a furcocercaria with morphological characteristics of Cercaria caratinguensis, and by an unknown furcocercaria. Drepanotrema cimex was infected by a furcocercaria with characteristics of C. caratinguensis and by C. macrogranulosa. No natural infection was found in B. oligoza. B. tenagophila showed no susceptibility to the experimental infection by the BH-MG strain of Schistosoma mansoni from Belo Horizonte and maintained at laboratory in B. glabrata snails.

Multicentric pheochromocytoma and involvement of the inferior vena cava.

CONTEXT: Extension of pheochromocytomas to the inferior vena cava is rare. Multicentric tumors are rare as well, being present in up to 10% of cases. Surgery is the treatment of choice because of the long-term survival free of disease. DESIGN: Case report. CASE REPORT: We report on a case of right adrenal pheochromocytoma with extension to the supra-diaphragmatic vena cava, which underwent surgical excision through thoracophrenic laparotomy without the need for cardiopulmonary bypass. In a 6-year follow-up, another pheochromocytoma was found in the infra-renal Zuckerkandl's organ. Complete surgical excision of the tumor was performed by a median laparotomy and complete retroperitoneal dissection. In both cases, the total removal of the pheochromocytoma has been guaranteed by having margins free of tumor and a normal post-operative level of catecholamines. The pathological study revealed a malignant pheochromocytoma with margins free of neoplasia in both specimens.

[Causes and prognosis of acute renal failure in elderly patients]. / Causas e prognóstico da insuficiência renal aguda hospitalar em pacientes idosos.

UNLABELLED: The prolonged life span of populations is the obvious reason for an increasing proportion of elderly patients with acute renal failure (ARF). The role of age as a factor indicative of a poor prognosis is a matter of controversy. OBJECTIVE: To evaluate this role we have analyzed the final outcome of elderly patients with ARF treated in our Nephrology Service. MATERIAL AND METHODS: Among 361 ARF cases prospectively studied during a two-year period (January 1995 to December 1996), 130 (36%) occurred in patients over 70 years of age. Etiology, clinical course and prognosis were analyzed. The average age was 76.0 +/- 4.7 years, varying from 70 to 94 years; 84 patients (65%) were male, and surgical causes accounted for 51% of geriatric ARF. RESULTS: The most frequent causes were: ischemic (volume depletion, arterial hypotension, and/or low cardiac output) in 48 patients (38%), sepsis in 40 (312%), nephrotoxic drugs in 46 (35%) and obstructive abnormalities in 10 (7.7%); in other 14 (11%) they were diagnosed more than a causal agent. Oliguria was present in 37. 2% (81 patients), and dialysis was needed in 50 patients (39%). The mortality in the population with age below 70 years was 43% and in the elderly patients was 53.8%; total mortality was similar in both groups of patients (p=0.085). Oliguria, need for dialysis, presence of surgical causes of ARF, and ARF acquired within the intensive care unit were associated with poor prognosis in elderly group. CONCLUSION: We concluded that the percentile of elderly patients with ARF is elevated; they presented mortality about of 50%, and this is not superior to the observed in the youngest population; oliguria, dialysis need, ICU cares and surgery are factors of unfavorable prognostic in these patients.

Human proximal femur bone adaptation to variations in hip geometry.

The study of bone mass distribution at proximal femur may contribute to understand the role of hip geometry on hip fracture risk. We examined how bone mineral density (BMD) of proximal femur adapts to inter individual variations in the femoral neck length (FNL), femoral neck width (FNW) and neck shaft angle (NSA). A parameterized and dimensionally scalable 3-D finite element model of a reference proximal femur geometry was incrementally adjusted to adopt physiological ranges at FNL (3.90-6.90cm), FNW (2.90-3.46cm), and NSA (109-141º), yielding a set of femora with different geometries. The bone mass distribution for each femur was obtained with a suitable bone remodelling model. The BMDs at the integral femoral neck (FN) and at the intertrochanteric (ITR) region, as well as the BMD ratio of inferomedial to superolateral (IM:SL) regions of FN and BMD ratio of FN:ITR were used to represent bone mass distribution. Results revealed that longer FNLs present greater BMD (g/cm(3)) at the FN, mainly at the SL region, and at the ITR region. Wider FNs were associated with reduced BMD at the FN, particularly at the SL region, and at the ITR region. Larger NSAs up to 129° were associated with BMD diminutions at the FN and ITR regions and with increases of the IM:SL BMD ratio while NSAs larger than 129° resulted in decrease of the IM:SL BMD ratio. These findings suggest hip geometry as moderator of the mechanical loading influence on bone mass distribution at proximal femur with higher FNL favoring the BMD of FN and ITR regions and greater FNW and NSA having the opposite effect. Augmented values of FNL and FNW seem also to favor more the BMD at the superolateral than at the inferomedial FN region.

Genotoxic evaluation, secondary metabolites and antioxidant capacity of leaves and roots of Urera baccifera Gaudich (Urticaceae).

In addition to phenolics, flavonoids, flavonols, alkaloids and condensed tannins, our tests identified the antioxidant and genotoxic properties in the crude extract (CE) and fractions of Urera baccifera (Urticaceae) roots and leaves. Oxalic acid (OA) content was determined by HPLC-DAD, which presented high values in the roots (1.82 ± 0.21, 1.79 ± 0.22 and 1.38 ± 0.15 mg/g in butanolic, CE and ethyl acetate fraction, respectively). OA caused a 30.7% reduction in the leucocyte proliferation, followed by butanolic fractions of roots (24.15%) and leaves (23.28%). The mitotic index was lower in butanolic fractions of leaves (8.7%) and roots (8.3%), similar to the OA index, which was 6.0%. The DNA damage index in cultured leukocytes was observed for OA (19.33) and butanol fraction treatments (22.67 and 16, respectively, for leaves and roots). Antioxidant capacity (DPPH and TBARS) was moderated, which was confirmed by the low phenolic, flavonol and flavonoid contents in both parts of the plant.

Feeding preference of the shredder Phylloicus sp. for plant leaves of Chrysophyllum oliviforme or Miconia chartacea after conditioning in streams from different biomes / Preferência alimentar do fragmentador Phylloicus sp. por folhas de Chrysophyllum oliviforme e Miconia chartacea após condicionadas em riachos de dois biomas

Abstract Macroinvertebrate shredders consume preferably leaves conditioned by fungi and bacteria which offer greater palatability to them. Plant species in Cerrado present high concentration of chemical elements such as lignin and cellulose, phenols and tanins thus making them less attractive for shredders consumption and limiting the palatability. This study aimed to evaluate the feeding preference of a macroinvertebrate shredder of the genus Phylloicus for plant material from two different biomes (Cerrado and Mata Atlântica), after conditioning in a stream of Mata Atlântica and observing their physical and chemical characteristics. Senescent leaves were collected, monthly from the litterfall of riparian vegetation in a 500 m stretch of a stream in each biome from August 2014 to January 201. The most abundant species in each stream was selected for the experiment. The experimental design consisted in with two treatments. The first (T1) comprised leaf discs from Chrysophyllum oliviforme (Cerrado species) together with leaf discs of Miconia chartacea (Atlantic Forest species) which were conditioned in the Atlantic Forest stream. The second treatment (T2) involved leaf discs of Miconia chartacea conditioned in Mata Altlântica and Cerrado streams. Both tests had showed significant differences between the two treatments (T1 and T2). For T1, there was consumption of M. chartacea leaf discs by Phylloicus sp., but there was no consumption of C. oliviforme discs. For T2, there was preference for M. chartaceae leaves conditioned in a stream of Mata Atlântica than in Cerrado stream. The results showed that Phylloicus sp., had presented preference for food detritus of the Mata Antlântica biome and rejection to the one from Cerrado biome.

Resumo Macroinvertebrados fragmentadores consomem folhas preferencialmente condicionadas por fungos e bactérias que lhes oferece uma maior palatabilidade. Nas espécies do cerrado esse condicionamento está também associado às altas concentrações de elementos químicos limitantes à palatabilidade como alto teor de lignina e celulose, que tornam as folhas menos atrativas para os fragmentadores. O trabalho teve como objetivo avaliar a preferência alimentar de macroinvertebrados fragmentador ( Phylloicus sp.) por material vegetal de dois diferentes biomas (Cerrado e Mata Atlântica), após condicionamento em riacho de Mata Atlântica, observando suas características físicas e químicas. Foram coletadas folhas senescentes do aporte vegetal (AV) de espécies nativas da vegetação ripária nesses dois biomas, com periodicidade mensal em um trecho de 500 m de um córrego em cada bioma. O experimento foi delineado com dois tratamentos. O primeiro (T1) compreendeu discos de folhas do Cerrado (Chrysophyllum oliviforme) mais discos de folhas da Mata Atlântica (Miconia chartacea) que foram condicionadas em córrego de Mata Atlântica. O segundo tratamento (T2) envolveu discos de folhas da Mata Atlântica condicionadas em córrego da Mata Atlântica mais discos de folhas da Mata Atlântica condicionadas em córrego do Cerrado. Os dois testes apontaram diferenças significativas entre os dois tratamentos (T1 e T2). Para T1 houve consumo de discos de folha de M. chartacea por Phylloicus sp, mas não houve consumo dos discos de C. oliviforme , de Cerrado. Para T2, houve o consumo, porém a preferência pelas folhas de M. chartaceae condicionadas no córrego da Mata Atlantica foi consideravelmete maior. Os resultados apontam que Phylloicus sp, apresentou preferência alimentar pelo detrito de bioma de Mata Atlântica e rejeição pelo detrito do bioma Cerrado.

Femoral neck bone adaptation to weight-bearing physical activity by computational analysis.

Individual differences in bone mass distribution at the proximal femur may be determined by daily weight-bearing physical activity (PA) since bone self-adapts according to the mechanical loads that is submitted. The aim of this study was to analyse computationally the effect of different weight-bearing PA types in the adaptation of the femoral neck (FN) by analysing regional differences in bone mineral density (BMD) at the integral FN and its superior, inferior, anterior and posterior subregions. To achieve this, it was adopted a 3-D femoral finite element (FE) model coupled with a suitable bone remodeling model. Different PA types were determined based both on ordinary lifestyle and mechanically more demanding PA as low magnitude impacts (L-I), moderate-magnitude impacts from odd directions (O-I) and high-magnitude vertical impacts (H-I). It was observed that as time spent in weight-bearing PA increases, BMD augment around the integral FN, but with different bone mass gain rates between subregions depending on the magnitude and directions of the hip contact forces; H-I was the type of weight-bearing PA which structurally most favor the gain of bone mass superiorly at the FN while both the H-I and the O-I types of PA promoted the largest bone mass gain rates at the anterior and posterior subregions of the FN. Because these types of weight-bearing PA were associated with a more uniform bone mass spatial distribution at the FN, they should provide a potential basis for targeted PA-based intervention programs for improving hip strength.

Composition of the essential oil of Buddleja thyrsoides Lam. leaves from Brazil.

This study is designed to examine the chemical composition of the essential oil obtained from Buddleja thyrsoides Lam leaves from south Brazil. Gas chromatography/mass spectrometry analysis resulted in the detection of 12 components, representing 91.4% of the total oil composition. Germacrene D (27.16%), 1,10-di-epi-Cubenol (13.37%), α-Cadinol (12.95%), Bicyclogermacrene (9.00%), Globulol (8.59%) and caryophylene E (5.26%) were the main components. To the best of our knowledge, this is the first study of the composition of essential oil of the B. thyrsoides collected in Brazil.

The protective effects of guaraná extract (Paullinia cupana) on fibroblast NIH-3T3 cells exposed to sodium nitroprusside.

The antioxidant effects of the hydro-alcoholic guaraná extract (Paullinia cupana var. sorbilis Mart.) on nitric oxide (NO) and other compounds generated from the degradation of sodium nitroprusside (SNP) in an embryonic fibroblast culture (NIH-3T3 cells) were evaluated. The guaraná bioactive compounds were initially determined by high-performance liquid chromatography: caffeine=12.240 mg/g, theobromine=6.733 mg/g and total catechins=4.336 mg/g. Cells were exposed to 10 µM SNP during a 6 h period because the cells exhibited >90% mortality at this concentration. Guaraná was added to the cultures in five concentrations (0.5, 1, 5, 10 and 20 mg/mL). The guaraná antioxidant effect was evaluated by viability assays, biochemical oxidation [lipid peroxidation, catalase and superoxide dismutase (SOD) activity] and genotoxicity (DNA Comet assay) analysis. Additionally, oxidative stress was evaluated by a 2,7-dihydrodichlorofluorescein diacetate fluorescence assay. Guaraná reverted the SNP toxicity mainly at lower concentrations (<5 mg), which decreased cell mortality, lipid peroxidation, DNA damage and cell oxidative stress as well as increased the SOD levels. These results demonstrate that guaraná has an antioxidant effect on NO metabolism in situations with higher cellular NO levels.

Association between interleukin-1 beta polymorphism (+3953) and obesity.

It now appears that obesity is associated with a low-grade inflammation of white adipose tissue resulting from chronic activation of the innate immune system as interleukin-1 beta (IL-1). Previous investigations have described a positive association between IL-1 beta +3953 (C>T) gene polymorphism (rs 1143634) and obesity, suggesting functional effects on fat mass, fat metabolism and body mass. However, it is necessary to determine if these results occur in other populations and if they are influenced by sex and age. Therefore, we performed a case-control study using 880 Caucasian subjects (59.7+/-11.9 years old) from the Brazilian Aging Research Program (non-overweight=283, overweight=334, obese=263) previously investigated in genetic studies, in whom we analyzed the IL-1 beta +3953C/T polymorphism. We observed higher T allele (CT/TT) frequency in non-overweight than overweight and obese groups. The odds ratio showed 1.340 (95% CI: 1.119-1.605) times more chance of the obese group being CC carriers compared to non-overweight group independent of sex and age. This study corroborates the idea that the IL-1 system is linked to the development of obesity.