8-hydroxyquinoline and quinazoline derivatives as potential new alternatives to combat Candida spp. biofilm.

Often associated to the colonization by Candida spp. biofilm, the catheter-related infections are a serious health problem since the absence of a specific therapy. Hence, the main objective of this work was to evaluate the activity of 8-hydroxyquinoline and quinazoline derivatives on Candida spp. biofilms. A quinazoline derivative (PH100) and an 8-hydroxyquinoline derivative (PH157) were tested against nine strains of C. albicans, C. tropicalis and C. parapsilosis, and their biofilms in polystyrene microtitre plates and on polyurethane central venous catheter. The PH157 compound was incorporated into a film-forming system-type formulation and its capacity to inhibit biofilm formation on catheters was evaluated. The compounds were active against planktonic and sessile cells, as well as against the tested biofilms. PH157 compound performed better than the PH100 compound. The formulation containing PH157 presented results very similar to those of the compound in solution, which indicates that its activity was preserved. Both compounds showed activity against Candida spp. strains and their biofilm, with better PH157 activity. The formulation preserved the action of the PH157 compound, in addition, it facilitates its application on the catheter. The structural modifications that these compounds allow can generate compounds that are even more active, both against planktonic cells and biofilms.

Effect of different dietary arachidonic, eicosapentaenoic, and docosahexaenoic acid content on selected immune parameters in gilthead sea bream (Sparus aurata).

The increasing replacement of fish oil (FO) by vegetable oils (VO) in aquafeeds for marine carnivorous species may lead to modifications of the dietary arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) levels and ratios that may compromise the fish immune status. Therefore, this study aimed to assess how different dietary ratios of ARA, EPA, and DHA modulate the immune response of gilthead sea bream juveniles. For that purpose, four isoproteic and isolipidic plant-based diets were formulated to include different dietary Long-Chain-Polyunsaturated Fatty Acids (LC-PUFA) ratios (%DM): Diet A (2.0% ARA: 0.2% EPA: 0.1% DHA); Diet B (1.0% ARA: 0.4% EPA: 0.4% DHA); Diet C (0% ARA: 0.6% EPA: 0.6% DHA); Diet D (0% ARA: 0.3% EPA: 1.5% DHA). Fish were fed the experimental diets for 56 days and then the haematological profile, humoral innate immune parameters, and expression of immune-related genes in the gut were determined. Our results showed that fish fed diet B exhibited an increase in the monocyte numbers, alternative complement activity, and bactericidal activity against Photobacterium damselae compared to the values found in fish fed other diets. Contrarily, peroxidase, antiproteases, and proteases activity and nitric oxide levels were not affected by the dietary treatments. Immune-related gene expression in the distal intestine did not show differences between dietary treatments. Overall, a diet with a balanced n-6 (ARA) and n-3 (EPA+DHA) LC-PUFA ratio seemed to better modulate the fish innate immune response, thus better preparing the fish innate immune system against potential immunological insults.

Oxidative status and intestinal health of gilthead sea bream (Sparus aurata) juveniles fed diets with different ARA/EPA/DHA ratios.

The present work assessed the effects of dietary ratios of essential fatty acids, arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic acid (DHA), on liver and intestine oxidative status, intestinal histomorphology and gut microbiota of gilthead sea bream. Four isoproteic and isolipidic plant-based diets were formulated containing a vegetable oil blend as the main lipid source. Diets were supplemented with ARA/EPA/DHA levels (%DM) equivalent to: 2%:0.2%:0.1% (Diet A); 1.0%:0.4%:0.4% (Diet B); 0%:0.6%:0.6% (Diet C); 0%:0.3%:1.5% (Diet D) and tested in triplicate groups for 56 days. Lipid peroxidation was higher in fish fed diets C and D while no differences were reported between diets regarding total, oxidized, and reduced glutathione, and oxidative stress index. Glutathione reductase was higher in fish fed diet A than diets C and D. No histological alterations were observed in the distal intestine. Lower microbiota diversity was observed in intestinal mucosa of fish fed diet C than A, while diets C and D enabled the proliferation of health-promoting bacteria from Bacteroidetes phylum (Asinibacterium sp.) and the absence of pathogenic species like Edwardsiella tarda. Overall, results suggest that a balance between dietary ARA/EPA + DHA promotes gilthead sea bream juveniles' health however higher dietary content of n-3 LC-PUFA might limited the presence of microbial pathogens in intestinal mucosa.

Ex vivo nail infection as an effective preclinical method for screening of new topical antifungals.

Onychomycosis are fungal nail infections comprising of about 50% of onychopathies and are commonly caused by dermatophytes. The treatment of this dermatomycosis requires a long period of time and is associated with high rates of recurrence. In view of the need to evaluate the antifungal performance of promising preclinical compounds, we developed, in this study, a practical and accessibleex vivo model for establishing a Trichophyton rubrum onychomycosis framework using porcine hooves. This model has as its main advantage the similar structural and three-dimensional characteristics that the porcine hooves have with the human nail. The proposed model allowed to evaluate the antifungal activity of a new antifungal compound and a reference drug (terbinafine), both already incorporated into a nail lacquer for topical use. Treatments with compound 3-selenocyanate-indole (Se4a) and with terbinafine incorporated into this nail lacquer completely inhibited fungal growth, corresponding to the profile of in vitro activity observed against T. rubrum. This study concludes that the ex vivo porcine hoof model is an effective alternative method for preclinical screening of drugs or new topical compounds developed to combat onychomycosis. Further studies are needed to compare the permeability of porcine hooves with human nails permeability.

A multiple endpoint approach reveals potential in vitro anticancer properties of thymoquinone in human renal carcinoma cells.

Thymoquinone (TQ) is a monoterpene isolated from the oil of Nigella sativa seeds. The aim of this work was to evaluate the cytotoxic effects induced by TQ and its impact on the migration and invasion potential of 786-O human renal cancer cells. These cells were exposed to TQ (1-100 µM) for 24 and 48 h and cell viability assessed using the Crystal Violet and MTS assays. TQ treatment clearly decreased cell viability in a concentration- and time-dependent manner. TQ exposure moderately increased intracellular ROS levels and co-incubation with reduced glutathione markedly increased cell viability. Moreover, the effect of TQ in the cell cycle distribution was evaluated using flow cytometry, and an increase in the sub-G1 population was observed, especially at 30 µM, along with an increase in the % of apoptotic cells. TQ did not show genotoxic effects at a non-cytotoxic concentration (1.0 µM). At this concentration level, TQ significantly decreased the collective migration of 786-O cells, whereas it had no effect in chemotactic migration. TQ also decreased the invasiveness potential of 786-O cells, as evaluated by the transwell invasion assay. Overall, these results suggest that TQ presents an anticancer potential in the context of renal cancer, warranting further investigation.

Long term prognosis of acute pulmonary embolism.

BACKGROUND: Acute pulmonary embolism (PE) can be fatal if left untreated. Long-term prognosis of acute PE in the 21st century has not been fully reported. We aimed to determine the long-term prognosis of patients hospitalized with acute PE and compare survivalof patients with idiopathic and secondary PE. MATERIALS AND METHODS: We retrospectively analysed a cohort of hospitalized patients with acute PE between 2006 and 2013. EXCLUSION CRITERIA: <18 years, venous embolism of non-pulmonary veins, chronic thromboembolic pulmonary hypertension, and presumptive diagnosis without image confirmation. Only patients with a first PE episode were included. End-point: all-cause mortality. Patients were compared according to PE aetiology: idiopathic, secondary to neoplastic conditions and secondary to non-neoplastic conditions. A Cox-regression analysis was used to study the prognostic impact of PE aetiology. RESULTS: We studied 872 hospitalized acute PE patients. Median age 70 years, 56.9% were women. PE was idiopathic in 376 (43.1%), secondary to a neoplastic condition in 284 (32.6%) and secondary to a condition other than neoplasia in 212 (24.3%). Patients were followed for a median 25 months period and 508 (58.3%) died. Patients with PE attributed to a neoplastic condition had the worst survival. Patients with idiopathic PE had a multivariate-adjusted HR of mortality of 1.46 (1.08-1.99) during the over 2-year follow-up period when compared to those with acute PE attributed to a non-neoplastic condition. CONCLUSIONS: Patients with idiopathic acute PE have an almost 50% higher death risk in a median 2-year follow-up period than those with acute PE secondary to a condition other than neoplasia.

Solid lipid nanoparticles containing copaiba oil and allantoin: development and role of nanoencapsulation on the antifungal activity.

The aim of this work was to develop solid lipid nanoparticles (SLN) containing copaiba oil with and without allantoin (NCOA, NCO, respectively) and to evaluate their antifungal activity. Nanoparticle suspensions were prepared using a high homogenisation technique and characterised by dynamic light scattering, laser diffraction, nanoparticle tracking analysis, multiple light scattering analysis, high-pressure liquid chromatography, pH and rheology. The antifungal activities of the formulations were tested in vitro against the emergent yeasts Candida krusei and Candida parapsilosis, and the fungal pathogens of human skin Trichophyton rubrum and Microsporum canis. The dynamic light scattering analysis showed z-average diameters (intensity) between 118.63 ± 8.89 nm for the nanoparticles with both copaiba oil and allantoin and 126.06 ± 9.84nm for the nanoparticles with just copaiba oil. The D[4,3] determined by laser diffraction showed similar results of 123 ± 1.73 nm for the nanoparticles with copaiba oil and allantoin and 130 ± 3.6 nm for the nanoparticles with copaiba oil alone. Nanoparticle tracking analysis demonstrated that both suspensions had monomodal profiles and consequently, the nanoparticle populations were homogeneous. This analysis also corroborated the results of dynamic light scattering and laser diffraction, exhibiting a smaller mean diameter for the nanoparticles with copaiba oil and allantoin (143 nm) than for the nanoparticles with copaiba oil (204 nm). The physicochemical properties indicated that the dispersions were stable overtime. Rheology evidenced Newtonian behaviour for both suspensions. Antifungal susceptibility showed a MIC90 of 125 µg/mL (nanoparticles with copaiba oil) and 7.8 µg/mL (nanoparticles with copaiba oil and allantoin) against C. parapsilosis. The nanoparticles with copaiba oil and the nanoparticles with copaiba oil and allantoin presented a MIC90 of 500 µg/mL and 250 µg/mL, respectively, against C. krusei. The MIC90 values were 500 µg/mL (nanoparticles with copaiba oil) and 1.95 µg/mL (nanoparticles with copaiba oil and allantoin) against T. rubrum. Against M. canis, the nanoparticles with copaiba oil and allantoin had a MIC9 of 1.95 µg/mL. In conclusion, nanoencapsulation improved the antifungal activity of copaiba oil, which was enhanced by the presence of allantoin. The MICs obtained are comparable to those of commercial products and can represent promising therapeutics for cutaneous infections caused by yeasts and dermatophytes.

Evaluation of physico-chemical characteristics of fresh, refrigerated and frozen Lacaune ewes' milk / Avaliação das características físico-químicas de leite fresco, resfriado e congelado de ovelhas Lacaune

The production of ewe milk is seasonal and milk yield per animal is low, even in specialized animals. This study aimed to verify the possibility of preserving bulk tank milk for seven days under cooling (5°C) and freezing (-5°C), verify the influence of cooling treatments and of the months of the year on the physical and chemical characteristics of the product. The chemical composition of milk, including the fat, protein, lactose and total solids contents, was not altered by cooling and freezing. Protein and lactose contents varied according to the months of the year. The average percentage and standard deviation of fat, protein, lactose and total solids was 8.10±1.30, 5.22±0.37, 4.43±0.23 and 19.34±1.54, respectively. The density, pH, titratable acidity, as well as alcohol and heat stability tests were significantly influenced by the treatments used (P < 0.05), but no differences were found between fresh and frozen milk. Prolonged refrigeration caused an increase in acidity and decrease in pH, with a consequent reduction in the stability of milk. These results demonstrated that freezing does not affect the chemical composition and physical characteristics of milk in nature and it could be a solution for the producer and the sheep milk industry...

O leite ovino, mesmo no caso de raças especializadas, apresenta sazonalidade de produção e baixa produtividade por fêmea. Em função disso, o objetivo do presente estudo foi verificar a possibilidade de conservação do leite fresco, por um período de sete dias, sob refrigeração e congelamento, verificando a influência dos tratamentos aplicados, e dos meses do ano, sobre as características físico-químicas do produto. A composição química, incluindo gordura, proteína, lactose e sólidos totais, não sofreu alterações com o resfriamento e congelamento do leite, contudo os teores de proteína e lactose variaram durante os meses do ano. O percentual médio de gordura, proteína, lactose e sólidos totais foi de 8,10±1,30, 5,22±0,37, 4,43±0,23 e 19,34±1,54, respectivamente. A densidade, o pH, a acidez titulável e as provas de estabilidade ao álcool e ao calor sofreram influência significativa dos tratamentos aplicados (P<0,05), não havendo diferenças entre o leite fresco e o leite congelado. O resfriamento prolongado causou aumento da acidez e diminuição do pH, com a consequente diminuição da estabilidade do leite. Os resultados do presente estudo demonstram que o congelamento não afeta a composição química e as características físicas do leite in natura, podendo ser uma solução para o produtor e para a indústria de leite ovino...

Influence of nanoencapsulation on the sensory properties of cosmetic formulations containing lipoic acid.

Sensory analysis has become a valuable tool in qualifying consumer perception regarding cosmetic products. This study aims to explore the application of discriminative and affective consumers sensory analysis in evaluating the influence of nanoencapsulation on the sensory properties and rheological characteristics of a cosmetic formulation containing lipoic acid. The nanocapsules were prepared by the pre-formed polymer precipitation method. Semisolid formulations were prepared using a silicone emulsion system, and these pseudoplastic fluids were characterized using rheological methods. The panellists (n = 88) analysed the formulations with and without nanoencapsulated lipoic acid as paired comparisons within the discriminative and affective sensory analysis. In these measurements, spreadability, stickiness, oiliness and sulphur odour were evaluated. The panellists had no previous training in conducting these measurements. It was shown that nanotechnology can change some sensory characteristics of the formulations. The spreadability, stickiness and oiliness are the attributes for which the panellists noted differences with statistical significance. The spreadability difference could be due to the results found in the rheological profiles and consistency indexes between formulations. In the discriminative analysis, the panellists could not detect any noticeable differences in the sulphur odour or residual properties between samples, attributes that would influence whether consumers adhered to the selected treatment. Considering affective analysis, the consumers communicated that the formulation containing the nanoencapsulated lipoic acid, which presented less consistency, was preferred based on the reduction in immediate stickiness and residual sulphur odour. The free lipoic acid formulation was preferred in terms of residual oiliness and spreadability.

Effects of temperature and dietary protein level on hepatic oxidative status of Senegalese sole juveniles (Solea senegalensis).

Effects of 55 and 45% dietary protein levels (55P and 45P diets, respectively) and temperature (12 and 18 °C) on hepatic activity of superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione reductase (GR), glucose-6-phosphate dehydrogenase and lipid peroxidation (LPO) levels of Solea senegalensis juveniles were studied. Further, effects of acute thermal shocks provoked by a drop (18 °C to 12 °C) or a rise (12 °C to 18 °C) of water temperature on sole oxidative state was also evaluated. Dietary protein reduction increased LPO levels though no major alterations were found on antioxidant enzyme activities between dietary treatments. At 12 °C GR activity was higher and SOD activity was lower than 18 °C but LPO levels were not affected. In both thermal shock cases, LPO levels increased in 55P group, probably due to insufficient antioxidant enzyme activation. In contrast, fish of 45P group under acute exposition to warmer and colder temperature exhibited no substantial changes and a significant decrease on LPO levels, respectively, along with no major changes in antioxidant enzymes. Overall, results suggest that independently of rearing temperatures 45P group was more susceptible to oxidative stress than 55P group. Thermal shock either due to rise or drop of temperature seemed to induce oxidative stress in 55P group.

Endometrial biopsy with uretral plastic No. 12 sonde.

The authors demonstrate technique for obtaining endometrial fragments in 25 patients sent to Gynecological Cancer Preventive Service of the Arnaldo Vieira de Carvalho Cancer Institute, during the period of September 1991 through January 1992, presenting abnormal uterine hemorrhage. Technique consists in vacuum aspiration with the support of a No. 12 uretral sonde, provided with one lateral opening only. Correlation with curettage was positive in 88% of the patients; biopsy with sonde did not result in confirmation in three patients only. No complications at all were observed (infection, hemorrhage or uterus perforation). Authors understand that this method is in conformity with a correct diagnostic performance, being a low cost procedure, almost painless, easy to be done and furnishing sufficient material for the histo-pathologic diagnosis, in 88% of the cases studied.