Application of biophysical and skin imaging techniques to evaluate the film-forming effect of cosmetic formulations.

OBJECTIVE: Products with film-forming effect, or 'second skin', which guarantees an immediate protective effect after application, is a highlight, especially when composed of natural ingredients. Thus, the objective of this study was to evaluate the immediate film-forming effect on skin of a gel and emulsion formulations added with Kappaphycus alvarezii and Caesalpinia spinosa extracts through biophysical and skin imaging techniques, especially with the Reflectance Confocal Microscopy (RCM). METHODS: The measurements were done in the forearm region before (baseline) and 1 h after of application of the developed formulation and its control. The parameters related to the stratum corneum water content, transepidermal water loss (TEWL), cutaneous microrelief and morphological and structural characteristics of the epidermis were analysed through the following biophysical and skin imaging techniques: Corneometer® CM 825, Tewameter® TM 300, Visioscan® VC98 and Vivascope® 1500, respectively. A sensorial analysis was also performed to study how the formulations were perceived on the skin. RESULTS: The obtained results showed that the active ingredient under study allows the film formation on the skin surface, leading to a reduction of TEWL and skin desquamation. The obtained images from RCM showed a reduction of furrows on the skin surface and a film formation after a single application of the formulations. However, these effects were more pronounced in the emulsion formulation, which suggests a synergistic effect of the active ingredient under study with the emollients of formulation composition. This result was also observed in the sensorial analysis, as both formulations added with the active substance were well evaluated. CONCLUSION: The presence of Kappaphycus alvarezii and Caesalpinia spinosa extracts in the studied cosmetic formulations, enabled a film formation on a skin surface, bringing benefits as a reduction of transepidermal water loss and skin desquamation, as well as a furrows reduction and an improvement of stratum corneum after 1 h of application. Finally, the skin imaging techniques can be suggested as an excellent tool to evaluate a film-forming effect of cosmetic formulations.

OBJECTIF: Les produits ayant un effet filmogène, ou « deuxième peau ¼ qui garantit un effet protecteur immédiat après l'application, sont un plus, notamment lorsqu'ils sont composés d'ingrédients naturels. L'objectif de cette étude visait ainsi à évaluer l'effet filmogène immédiat sur la peau de formulations avec gel et émulsion enrichies d'extraits de Kappaphycus alvarezii et de Caesalpinia spinosa grâce à des techniques d'imagerie cutanée et biophysique, en particulier par microscopie confocale par réflectance (MCR). MÉTHODES: Les mesures ont été effectuées dans la région de l'avant-bras avant (référence) et 1 h après l'application de la formulation développée et de son contrôle. Les paramètres liés à la teneur en eau de la couche cornée, perte d'eau transépidermique (Transepidermal Water Loss, TEWL), au microrelief cutané et aux caractéristiques structurelles et morphologiques de l'épiderme ont été analysés à l'aide des techniques d'imagerie cutanée et biophysique suivantes : Corneometer® CM 825, Tewameter® TM 300, Visioscan® VC98 et Vivascope® 1500, respectivement. Une analyse sensorielle a également été réalisée pour étudier comment les formulations étaient perçues sur la peau. RÉSULTATS: Les résultats obtenus ont montré que le principe actif étudié permet la formation d'un film à la surface de la peau, entraînant une réduction de la TEWL et de la desquamation de la peau. Les images obtenues par MCR ont montré une réduction des sillons à la surface de la peau et la formation d'un film après une seule application des formulations. Ces effets étaient toutefois davantage prononcés avec la formulation de type émulsion, ce qui suggère un effet synergique du principe actif étudié avec les émollients contenus dans la formulation. Ce résultat a également été observé dans l'analyse sensorielle, car les deux formulations enrichies en substance active ont été bien évaluées. CONCLUSION: La présence d'extraits de Kappaphycus alvarezii et de Caesalpinia spinosa dans les préparations cosmétiques étudiées a permis la formation d'un film à la surface de la peau, procurant des bénéfices sous forme d'une réduction de la perte d'eau transépidermique et de la desquamation de la peau, de même qu'une réduction des sillons et une amélioration de la couche cornée 1 h après l'application. Enfin, les techniques d'imagerie cutanée peuvent être proposées comme excellent outil d'évaluation de l'effet filmogène des préparations cosmétiques.

Characterization of oily mature skin by biophysical and skin imaging techniques.

BACKGROUND: The skin is a complex biological system and may suffer change according to the environmental factors, as higher temperatures can increase sebum excretion, presenting oiliness and acne. These alterations can persist during the aging and provoke more changes in aged skin. In this study we evaluated the mature oily skin characteristics using biophysical and skin imaging techniques. MATERIAL AND METHODS: Sixty healthy female subjects, aged between 39 and 55 years old were recruited and separated into 2 groups according to their skin type: normal/dry and oily skin. The skin was evaluated in terms of stratum corneum water content, transepidermal water loss (TEWL) sebum content, dermis thickness and echogenicity, skin microrelief, and pores content. RESULTS: The mature oily skin presented no significant differences when compared to the normal/dry skin on the stratum corneum water content and TEWL parameters. The sebum content was significantly higher on the oily skin group. The microrelief analysis showed an increase of skin roughness values in the oily skin and increase of scaliness in the normal/dry skin. The oily skin showed lower dermis echogenicity mainly in the frontal region and higher dermis thickness when compared to normal/dry skin. CONCLUSION: The mature oily skin showed different characteristics from normal/dry skin in terms of sebum content, microrelief parameters, and dermis thickness. This way, the characterization of mature oily skin in an objective way is very important to development of dermocosmetic products for more effective treatments focused specially on this type of skin.

Human predisposition to cognitive impairment and its relation with environmental exposure to potentially toxic elements.

New lines of evidence suggest that less than 10% of neurodegenerative diseases have a strict genetic aetiology and other factors may be prevalent. Environmental exposures to potentially toxic elements appear to be a risk factor for Parkinson's, Alzheimer's and sclerosis diseases. This study proposes a multidisciplinary approach combining neurosciences, psychology and environmental sciences while integrating socio-economic, neuropsychological, environmental and health data. We present the preliminary results of a neuropsychological assessment carried out in elderly residents of the industrial city of Estarreja. A battery of cognitive tests and a personal questionnaire were administered to the participants. Multivariate analysis and multiple linear regression analysis were used to identify potential relationships between the cognitive status of the participants and environmental exposure to potentially toxic elements. The results suggest a relationship between urinary PTEs levels and the incidence of cognitive disorders. They also point towards water consumption habits and profession as relevant factors of exposure. Linear regression models show that aluminium (R 2 = 38%), cadmium (R 2 = 11%) and zinc (R 2 = 6%) are good predictors of the scores of the Mini-Mental State Examination cognitive test. Median contents (µg/l) in groundwater are above admissible levels for drinking water for aluminium (371), iron (860), manganese (250), and zinc (305). While the World Health Organization does not provide health-based reference values for aluminium, results obtained from this study suggest that it may have an important role in the cognitive status of the elderly. Urine proved to be a suitable biomarker of exposure both to elements with low and high excretion rates.

Disseminated histoplasmosis and AIDS: a prospective and multicentre study to evaluate the performance of different diagnostic tests.

The burden of histoplasmosis has been poorly documented in most of the endemic areas for the disease, including Brazil. Also, modern non-culture-based diagnostic tests are often non-available in these regions. This was a prospective cohort study in HIV-infected patients with suspected disseminated disease evaluated with different diagnostic tests. Patients were enrolled in three referral medical centres in Porto Alegre, Brazil. Among 78 evaluated patients, disseminated histoplasmosis was confirmed in eight individuals (10.3%) by the means of classical (culture/histopathology) tests. Antigen detection in the urine was found to be more sensitive: IMMY® ALPHA ELISA detected 13 positive cases (16.7%) and the in-house ELISA test developed by the Centers for Disease Prevention and Control (CDC) detected 14 (17.9%). IMMY® and CDC tests provided concordant results in 96.2% of cases. This is the first study to compare the performance of the in-house CDC ELISA test with the IMMY® commercial test for the diagnosis of histoplasmosis, and a high degree of concordance was observed. The study revealed that H. capsulatum is an important agent of disseminated disease in AIDS patients in Brazil, reinforcing the importance of making available modern diagnostic tests as well as safer antifungal agents for the treatment of histoplasmosis.

Evaluation of antioxidant capacity of 13 plant extracts by three different methods: cluster analyses applied for selection of the natural extracts with higher antioxidant capacity to replace synthetic antioxidant in lamb burgers.

The aims of this study were: to evaluate the total equivalent antioxidant capacities (TEAC) and phenolic contents of 13 plants extracts; to select the most promising extracts regarding reducing activity using cluster analysis multivariate statistical technique; and to analyse evaluate sensory acceptance of lamb burgers produced with the most promising natural antioxidants replacing sodium erythorbate. Plant extracts were evaluated regarding TEAC by DPPH(•) and FRAP methods, and total phenolics contents by Folin-Ciocalteau assay. The TEAC values ranged from 0.50 to 9.06 g trolox/100 g dry weight (dw) and from 43.6 to 472.32 µmol trolox/g dw for DPPH(•) and FRAP methods, respectively, and the total phenolic contents from 5.98 to 74.01 mg GAE/g dw. Extracts from Origanum vulgare, Melissa officinalis, Origanum majorana L. and Rosmarinus officinalis were grouped as the ones with higher antioxidant capacities by cluster analysis. All burgers produced with each one of these four plant extracts or with sodium erythorbate showed no differences (P > 0.05) regarding consumers' sensory acceptance. In conclusion, it is possible to replace sodium erythorbate in lamb burgers by any of the four natural extracts selected without compromising sensory acceptance of this meat product.

Induction of oxidative stress in Prototheca zopfii by indole-3-acetic acid/HRP or 2,4-pentanedione/HRP systems and their oxidation products.

We investigated the toxic effects on Prototheca zopfii of indole-3-acetic acid (IAA) and 2,4-pentanedione (PD) combined with horseradish peroxidase (HRP) alongside the oxidation products of 3-methyl-2-oxindole (MOI) and indole-3-carbinol (I3C) from the IAA/HRP system and methylglyoxal (MGO) from the PD/HRP system. The microorganism was incubated in the absence (control) or presence of IAA, PD, IAA/HRP, PD/HRP, MOI, I3C and MGO and determined: (1) cytotoxicity by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay; (2) growth inhibitory concentration by resazurin assay and (3) antioxidant enzymes activities of: catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD). P. zopfii was more susceptible to IAA at 40 mM than PD at the same concentration, which seems to indicate that IAA was more effective at initiating cell death. These data corroborate results from the resazurin assay. Concentrations of 40 mM of IAA, IAA/HRP and PD/HRP, 20 mM of PD/HRP, 10 mM of MOI, 2 mM of I3C and 8 mM of MGO inhibited the growth of P. zopfii. With sub-inhibitory concentrations of IAA and IAA/HRP at 30 mM, MOI at 8 mM and I3C at 1 mM, the activities of CAT and GR increased, whereas no statistical difference was observed for CAT activity with IAA/HRP. Thus, PD at 30 mM and MGO at 6 mM increased the activities of CAT and GR, whereas PD/HRP system at 15 mM decreased CAT activity and PD/HRP and MGO showed no statistical difference for SOD activity. In conclusion, IAA/HRP or PD/HRP systems and their oxidation products exert cytotoxic effects on P. zopffi; however, I3C and MGO appear to exert greater microbicidal effect on P. zopfii.

B-cell epitopes of antigenic proteins in Leishmania infantum: an in silico analysis.

Serodiagnosis of visceral leishmaniasis is often hindered by cross-reactions to other parasitic diseases. Identifying specific B-cell epitopes in proteins is therefore important for immunodiagnostics, as well as for disease control by vaccines. This study aimed to identify linear and conformational B-cell epitopes and to evaluate the secondary structure of antigen proteins in Leishmania infantum using in silico analysis. Linear epitopes were predicted using the Immune Epitope Database and Analysis Resource (IEDB), BepiPred and BcePred programs. The conformational B-cell epitopes were identified using the CBTOPE server. The combination of the predictions using IEDB, BepiPred and BcePred generated 148 linear epitopes from the calpain-like cysteine peptidase (CP), thiol-dependent reductase 1 (TDR1) and HSP70 proteins. In total, 164 conformational epitopes were predicted, mostly located in the linear epitope region. The predicted epitopes are located in α helix and random coil regions in the thiol-dependent reductase 1 and HSP70 proteins. New linear and conformational B-cell epitopes of L. infantum proteins were identified in silico, and the prediction using various programs ensures greater accuracy of the results, as suggested by confirmation of previously identified HSP70 epitopes.

GSTT1, GSTM1, and GSTP1 polymorphisms as a prognostic factor in women with breast cancer.

The glutathione S-transferase (GST) family comprises phase-II cellular detoxification enzymes that catalyze the conjugation of chemotherapy drugs to glutathione and act on the apoptotic pathway. The aim of this study was to determine whether polymorphisms of the GSTT1, GSTM1, and GSTP1 genes are associated with different rates of overall survival (OS) and disease-free survival (DFS) after neoadjuvant chemotherapy in the management of locally advanced breast cancer, using either simple or combined analyses, and in relation to the post-therapy axillary lymph node status. Forty women with invasive ductal carcinoma of the breast submitted to neoadjuvant chemotherapy with 5-fluorouracil, epirubicin, and cyclophosphamide were genotyped for GSTT1, GSTM1, and GSTP1. Comparisons were performed for the three genes, either isolated or in pairs, in polymorphic or wild-type combinations. Finally, the OS and DFS of patients were analyzed with respect to axillary lymph node status and with respect to wild-type or polymorphic presentations of each gene. No statistically significant difference in OS and DFS was evident between women with wild-type or polymorphic forms of the genes, either isolated or in pairs, after neoadjuvant chemotherapy. By contrast, after treatment, lymph node-negative women had better OS and DFS only in the presence of polymorphisms of GSTP1, and improved DFS only in the presence of the polymorphic types of GSTT1 and GSTM1 compared to women with positive lymph nodes. The presence of polymorphic forms of GSTP1, GSTM1, and GSTT1 was crucial to conferring better OS and DFS among women with negative axillary lymph nodes.

Chemotherapy-induced oral mucositis: hierarchical analysis of recurrence factors.

PURPOSE: This study aimed to analyze, through a hierarchical model, the risk factors associated with the recurrence of chemo-induced oral mucositis (OM) in children and adolescents. METHODS: A retrospective cohort with 31 individuals of both sexes, aged 1-18 years, who were undergoing chemotherapy, and presented OM lesions was conducted. Data collection included analysis of medical records, interviews, and intraoral examination. Information regarding patients' socioeconomic and demographic profile, underlying disease, antineoplastic regimen, hematological condition, and oral health status were collected. To assess the association of independent variables with the outcome, the Chi-square, Fisher's Exact, and Mann-Whitney tests were used, in addition to a binary logistic regression model, with a maximum error of 5% and a 95% confidence interval. RESULTS: Significant associations were observed between the history of OM and the diagnosis of the child/adolescent, neutrophil count, previous cancer treatments and the chemotherapy scheme in use (p < 0.05). Binary logistic regression revealed a 13.69 higher risk of developing OM recurrence in individuals who received high-dose methotrexate (MTX) therapy. CONCLUSION: Socioeconomic and demographic factors did not influence OM recurrence. However, clinical variables, such as neutropenia, diagnosis of leukemia, and high-dose MTX protocols increase the chance of OM new cases.

Vaginal cone for postmenopausal women with stress urinary incontinence: randomized, controlled trial.

OBJECTIVE: To investigate the effect of vaginal cones and pelvic floor muscle training (PFTM) in postmenopausal women with stress urinary incontinence. METHODS: This randomized, controlled study included postmenopausal women, who complained of stress urinary incontinence. Forty-five women were allocated to three groups: a group given therapy with vaginal cones (n = 15), a group receiving therapy with PFTM (n = 15), and the control group (n = 15). Subjects in the intervention groups were treated for 6 weeks with twice-weekly sessions of 40 min. Women in the vaginal cone group carried out the pelvic floor muscle strengthening with vaginal cones. The control group did not receive any treatment during the corresponding time. They were evaluated before, at the end of treatment and 6 weeks after treatment completion for primary outcomes (1-h pad test for urinary loss and pelvic floor muscle pressure) and secondary outcomes (quality of life with King's Health Questionnaire, satisfaction with treatment, and continuity of training). RESULTS: For urinary leakage, there were statistical differences between the treated groups and the control group at the end of treatment and 6 weeks after treatment (all p < 0.01; effect size: vaginal cone group 20.97; PFMT group 20.96). The same behavior was shown for treatment with pelvic floor muscle pressure (all p < 0.01; effect size: vaginal cone group 22.58; PFMT group 21.68). There were no differences between the vaginal cone and PFMT groups in any of the evaluations. In outcomes for quality of life, significant differences were observed for incontinence impact and gravity domains when both treated groups were compared with the control group after treatment. Both groups reported similar satisfaction levels and the vaginal cone group demonstrated lower training continuity. CONCLUSION: Based on this study, there were similar positive results for treatment with the vaginal cone and pelvic floor muscle training for urinary leakage, pelvic floor muscle pressure and quality of life for postmenopausal women with stress urinary incontinence after 6 weeks.

Dopamine depletion in wistar rats with epilepsy.

The dopamine content in cerebral structures has been related to neuronal excitability and several approaches have been used to study this phenomenon during seizure vulnerability period. In the present work, we describe the effects of dopamine depletion after the administration of 6-hidroxidopamine (6-OHDA) into the substantia nigra pars compacta of male rats submitted to the pilocarpine model of epilepsy. Susceptibility to pilocarpine-induced status epilepticus (SE), as well as spontaneous and recurrent seizures (SRSs) frequency during the chronic period of the model were determined. Since the hippocampus is one of main structures in the development of this experimental model of epilepsy, the dopamine levels in this region were also determined after drug administration. In the first experiment, 62% (15/24) of 6-OHDA pre-treated rats and 45% (11/24) of those receiving ascorbic acid as control solution progressed to motor limbic seizures evolving to SE, after the administration of pilocarpine. Severeness of seizures during the model´s the acute period, was significantly higher in epileptic experimental rats (56.52%), than in controls (4.16%). In the second experiment, the frequency of seizures in the model's chronic phase did not significantly change between groups. Our data show that dopamine may play an important role on seizure severity in the pilo's model acute period, which seems to be due to dopamine inhibitory action on motor expression of seizure.

In vitro effect of children's toothpaste on brushing abrasion of eroded primary enamel.

PURPOSE: Emerging data have suggested that acid erosion has become an increasing clinical problem in pediatric dentistry. This study aimed to investigate the abrasive effects of two commercial toothpastes on primary enamel eroded by orange juice using an in vitro model. METHODS: Thirty enamel slabs were obtained from primary teeth and then randomly assigned to three groups (n = 8) comprising two different toothpastes: G1-control-distilled water; G2-Dentifrice A (containing no fluoride); and G3-Dentifrice B (1.100 ppmF-NaF). Each slab had one half protected to provide a control side, and the other one was subjected to treatments. The slabs were submitted to daily erosive challenges (3×/day, 2 min) in concentrated orange juice (pH 3.38) associated with abrasive challenges using a tooth-brushing machine (150 brush movements for each cycle). During the experimental period, the slabs were kept in 37 °C artificial saliva, and the experiment was carried out for 5 days. The depths of the resulting eroded areas were measured by stylus profilometry. The data were analyzed using ANOVA and the Tukey-Kramer test (α = 5%). RESULTS: Tooth loss (µm, mean ± SD) was 2.46 ± 1.18 for G1, 3.32 ± 2.12 for G2 and 2.14 ± 1.03 for G3. Therefore, the NaF dentifrice (Dentifrice B) produced significantly less mineral loss (p = 0.04) than dentifrice A. CONCLUSIONS: The findings suggest that fluoride toothpaste could protect primary enamel against erosion.

Apoptosis Induction in Murine Melanoma (B16F10) Cells by Mannosylerythritol Lipids-B; a Glycolipid Biosurfactant with Antitumoral Activities.

Mannosylerythritol lipids have drawn attention to cosmetic and pharmaceutical industries due to their non-toxicity and excellent biological interactions with human skin, particularly with the deepest epidermal layer. Lamellar liquid crystal structure, formed by MEL-B, is an interesting feature due to its similarity to the stratum corneum molecular arrangement and cell signaling events involved in the deregulation of the cancerous cell membrane. Thus, this work aimed to evaluate the cytotoxicity of commercial mannosylerythritol lipids-B in murine melanoma, fibroblast, and human erythrocytes cells. Cytotoxic effect was more pronounced on the tumor cells from 20 µg/mL, reducing cell viability by 65%, whereas fibroblast and human erythrocytes cells were more resistant to glycolipid treatment. Fluorescence microscopy and flow cytometer proved that mannosylerythritol lipids-B is an apoptosis inducer in tumor cells related to reactive oxygen species generation.

The effect of carbon and nitrogen sources on bovicin HC5 production by Streptococcus bovis HC5.

AIMS: To investigate the effect of media composition and agroindustrial residues on bovicin HC5 production by Streptococcus bovis HC5. METHODS AND RESULTS: Batch cultures of S. bovis HC5 were grown in basal medium containing different carbon and nitrogen sources. The activity of cell-free and cell-associated bovicin HC5 was determined in culture supernatants and acidic extracts obtained from cell pellets, respectively. Streptococcus bovis HC5 produced bovicin using a variety of carbon and nitrogen sources. The highest specific activity was obtained in media containing 16 g l(-1) of glucose, after 16 h of incubation. The peak in cell-free and cell-associated bovicin HC5 activity was detected when S. bovis HC5 cultures reached stationary phase. The bovicin HC5 specific activity and bacterial cell mass increased approximately 3-fold when yeast extract and trypticase (0.5 and 1.0 g l(-1), respectively) were added together to the basal medium. Streptococcus bovis HC5 cultures produced bovicin HC5 in cheese whey and sugar cane juice and maximal volumetric productivity was obtained after 12 h of incubation. CONCLUSIONS: Streptococcus bovis HC5 is a versatile lactic acid bacterium that can utilize several carbon and nitrogen sources for bovicin HC5 production. This bacterium could be a useful model to study bacteriocin production in the rumen ecosystem. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of agroindustrial residues as carbon sources could have an economical impact on bovicin HC5 production. To our knowledge, this is the first report to show the use of sugar cane juice for bacteriocin production by lactic acid bacteria.

Immigrants' access to health care: Problems identified in a high-risk tuberculosis population.

INTRODUCTION: Immigrants may utilize health care services differently than other residents and may also have a greater risk for tuberculosis (TB). OBJECTIVE: Identify barriers to healthcare access by immigrants, factors associated with these barriers, and discuss strategies that may reduce these barriers. MATERIAL AND METHODS: Anonymous questionnaires were given to immigrants at National Immigrant Support Centres between 2015 and 2016. Barriers to healthcare were identified using logistic regression. RESULTS: One-hundred and nineteen questionnaires were administered to immigrants, 9 of whom (8%) presented with TB while in Portugal. Twenty-one percent of immigrants reported barriers to healthcare access, and 69% had general practitioners (GPs). The presence of barriers to healthcare access was negatively associated with having a GP and with being married or in a de facto union. CONCLUSIONS: A considerable proportion of immigrants reported having difficulties accessing healthcare services in Portugal where legally these barriers are nonexistent. Certain factors were associated with these difficulties.

European case studies supporting the derivation of natural background levels and groundwater threshold values for the protection of dependent ecosystems and human health.

The EU Water Framework and Groundwater directives stipulate that EU member states have to assess groundwater chemical status by the use of groundwater threshold values derived for the protection of dependent ecosystems and human health. This paper presents a synthesis of main results of 14 European case studies evaluating a methodology for derivation of natural background levels (NBLs) and groundwater threshold values (TVs) proposed by the EU research project "BRIDGE". The 14 investigated groundwater bodies were selected to represent as many aquifer types, climate settings and European ecoregions as possible within the project group that included partners from 17 EU countries. The selected case study sites include transboundary groundwater bodies, EU Pilot River Basins and other important groundwater systems. Some are known to interact with associated ecosystems, while others do not. The proposed method derives groundwater threshold values based on environmental objectives for dependent ecosystems or groundwater "itself" using relevant reference criteria such as natural background levels, environmental quality standards and drinking water standards. Derived groundwater threshold values for dissolved Cl and As applying drinking water standards as examples of reference values, are compared for all 14 case studies. Additionally, more detailed analyses are conducted for the Odense Pilot River Basin and the Vouga River Basin, where groundwater threshold values for N and P and Cl, As, Cd, Ni, Pb, and Zn, respectively, are derived based on environmental objectives and quality standards for groundwater dependent ecosystems. Results demonstrate that the proposed methodology is operational and may be used to protect human health and the environment. Further they show that groundwater threshold values derived from environmental objectives and environmental quality standards for dependent ecosystems in some cases may be significantly lower than drinking water standards, e.g. for nitrate.

Assessment of the stability of sheep sausages with the addition of different concentrations of Origanum vulgare extract during storage.

The aim of this study was to assess the stability of sheep sausages with the addition of different concentrations of Origanum vulgare extract during storage. Five treatments were prepared: without natural antioxidant (control), sodium erythorbate 500mg/kg (ER), and three amounts of extract (N1=4964.51mg/kg, N2=6630.98mg/kg and N3=8038.20mg/kg). From appearance sensory analysis, control treatment differed significantly compared to ER (P<0.05) and N3 (P<0.01) groups, with intense red color, agreeing with trend of a* values. On the other hand, oregano extract improved the lipid and protein stability of cooked sausages during the storage time. Regarding volatile compounds from lipid oxidation, the N2 group presented the lowest values at the end of chilled period. In conclusion, the oregano extract showed antioxidant potential equivalent to sodium erythorbate at intermediate and high levels, calculated by DPPH∙ and FRAP methods, and most of the parameters were not compromised, allowing synthetic antioxidants replaced while maintaining the nutritional and sensory quality of cooked sausages.

Evaluation of oxidative stability of lamb burger with Origanum vulgare extract.

The objective was to evaluate replacement of sodium erythorbate with a natural antioxidant (oregano extract) on physicochemical and sensory stability of lamb burgers, and determine the appropriate amount. Five treatments were prepared, including control (without antioxidant), sodium erythorbate, and three concentrations of oregano extract (13.32, 17.79 and 24.01mL/kg), based on antioxidant capacity determined using the Folin-Ciocalteu, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) methods, respectively. Burgers containing oregano extract, at the concentration determined by FRAP method, had higher oxidative stability, evidenced by an 80% reduction (P<0.001) in thiobarbituric acid reactive substances, effective inhibition of protein oxidation (P<0.01) and less colour loss during frozen storage. Oregano extract did not impair (P>0.05) consumers' sensory acceptance of the lamb burgers. Under the conditions tested, addition of 24mL/kg of oregano extract could be recommended as a natural antioxidant in lamb burgers.

Effects of two diet techniques and delivery mode on weight loss, metabolic profile and food intake of obese adolescents: a fixed diet plan and a calorie-counting diet.

The aim of this study is to compare the weight loss of obese adolescents on two different low-calorie diets: fixed diet plan and calorie-counting diet. This is a randomized clinical study with 66 obese adolescents (body mass index Z score (ZBMI)>+3, 13.7±0.7 years, 60.6% male) with anthropometric, food intake, physical activity, laboratory, body composition and stage of pubertal development data evaluated. There was a reduction in the ZBMI in both groups (P<0.0001), without significant difference between them (P=0.87). There was a significant reduction in insulin, and homeostasis model assessment insulin resistance (HOMA-IR), with no difference between groups. A reduction in total energy intake of the groups was found, with an increase in the proportion of protein and reduction in carbohydrates. In this cohort of severely obese adolescents, fixed diet plan and calorie-counting diet led to a similar reduction of ZBMI, metabolic markers and total energy intake.