Nannochloropsis oceanica microalga feeding increases long-chain omega-3 polyunsaturated fatty acids in lamb meat.

To test the hypothesis that lambs fed freeze-dried Nannochloropsis oceanica (NO) biomass will have a higher deposition of EPA in tissues than those fed other Nannochloropsis EPA-sources, we fed 28 lambs with one of four diets: i) C, control, without EPA; ii) O, with 1.2% Nannochloropsis oil; iii) SD, with 12.3% spray-dried NO biomass; iv) FD, with 9.2% freeze-dried NO biomass. Dry matter intake, growth, tissues fatty acid composition, oxidative stability and sensory traits of the resultant meat were evaluated. The EPA was highest in tissues of lambs fed SD and FD compared with O but was similar between SD and FD. Total trans-18:1 did not differ among treatments, but the t10/t11-18:1 ratio decreased with all EPA containing diets. EPA diets were also supplemented with Vitamin E preventing the lipid oxidation in EPA-enriched meat and the meat sensory traits were not affected although occasionally some off-flavours were detected in FD meat.

MC3T3-E1 cell response to microporous tantalum oxide surfaces enriched with Ca, P and Mg.

The formation of a porous oxide surface doped with osteoconductive elements, Ca, P and Mg, to enhance osseointegration, was achieved through micro arc oxidation. Micro arc oxidation parameters, such as electrolyte composition, concentration and applied voltage, were studied to understand their effect on the morphology and chemical composition of the samples surface. Considering the optimum atomic concentration reported in literature for each osteoconductive element, microporous Ta anodic oxide samples treated with calcium acetate (CaA) and ß-glycerophosphate (ß-GP) revealed that an increase of ß-GP molarity in the electrolyte boosts Ca incorporation, as well as, increasing the porosity. In adding magnesium acetate (MgA) to the electrolyte, when composed by CaA + ß-GP, both addition and variation of MgA did not affect the surface morphology along the samples, being incorporated into the oxide layer for 0.1 M. Finally, in vitro tests were carried out to study the biocompatibility of Ta, to verify the cytotoxicity of the samples and their behavior towards cells, by performing adhesion and differentiation tests with the MC3T3-E1 cell line. Cytotoxicity tests revealed that the samples were non-toxic. Despite none of the samples having been raised up through cell adhesion tests, cell differentiation revealed promising results for the Ta-CaP.

Study of macro and microelements in fish from the Cienfuegos Bay. Relationship with its content in sediments.

Thirteen microelements (Al, As, Cd, Co, Cr, Cu, Fe, Li, Mn, Ni, Ti, V, and Zn) and four macroelements (Ca, K, Mg, and Na) were determined in the edible muscle tissue of fish in Cienfuegos Bay. Eight species, Albula vulpes, Diapterus rhombeus, Gerres cinereus, Haemulon carbonarium, Haemulon sciurus, Micropogonias furnieri, Kyphosus sectatrix, and Lutjanus cyanopterus were studied. The total concentrations and extracted concentrations using HCl were also determined in sediments. The quality of the analysis was guaranteed by validating the used analytical methods with certified reference materials of fish and sediments. The high content of macroelements confirmed the importance of fish as a valuable source of food for the local population. New information is provided on bioaccumulation of toxic elements As, Cd, Cu, Zn, and Cr in fish of the bay, with concentrations that overcame the local or international maximum allowable levels for human consumption in most samples analyzed. A high correlation between the content of toxic elements As, Cd, Zn, and macroelement K in fish and its concentration extracted from sediments using the HCl extraction method was observed.

A proteomic road to acquire an accurate serological diagnosis for human tegumentary leishmaniasis.

Diagnostic tools are important for clinical management and epidemiological evaluation of Tegumentary (TL) and Visceral (VL) Leishmaniasis. Serology is not frequently used for the diagnosis of the TL form because low antibody titers and cross-reaction with VL. Therefore, it is crucial to identify specific and immunogenic antigens from species associated with the TL form. Here we employed a proteomic approach coupled to an in silico analysis and identified the most abundant and immunogenic proteins from Leishmania amazonensis, Leishmania braziliensis and Leishmania infantum. Of 16 species specific proteins, nine were from the species causative of the TL form (L. amazonensis and L. braziliensis). In silico analysis revealed 18 B-cell epitopes with 0% similarity to Trypanosoma cruzi orthologs and, therefore, less likely to crossreact with sera of patients with Chagas disease. Two proteins reacted exclusively with serum from TL patients and presented several B-cell epitopes without similarity to T. cruzi orthologs: the hypothetical protein GI 134063939 and the metallo-peptidase Clan MA(E)-Family M3. The immunoassay revealed nine peptides with strong reactivity to sera from TL patients. These proteins and peptides may be good candidates to improve the specificity and sensibility of serological tests aiming to diagnose the TL of this neglected human disease. BIOLOGICAL SIGNIFICANCE: As no gold-standard test for tegumentary leishmaniasis (TL) exists, a combination of different diagnostic techniques is often necessary to obtain precise results. Thus, the identification of species-specific, highly immunogenic and abundant proteins that stimulate the humoral immune response in the host should help in the development of serological tests for human TL. Herein we searched for these potential antigens in Leishmania species related to American Leishmaniasis (L. amazonensis, L. braziliensis and L. infantum). To this end, we employed an immunoproteomic approach using proteins from these Leishmania species and sera from TL and Visceral Leishmaniasis (VL) patients. Our study unveils specific proteins and peptides that may represent antigens that will help the efforts to improve the accuracy of serological tests to diagnose the TL form.

Immunoproteomic and bioinformatic approaches to identify secreted Leishmania amazonensis, L. braziliensis, and L. infantum proteins with specific reactivity using canine serum.

Leishmania spp have a wide range of hosts, and each host can harbor several Leishmania species. Dogs, for example, are frequently infected by Leishmania infantum, where they constitute its main reservoir, but they also serve as hosts for L. braziliensis and L. amazonensis. Serological tests for antibody detection are valuable tools for diagnosis of L. infantum infection due to the high levels of antibodies induced, unlike what is observed in L. amazonensis and L. braziliensis infections. Likewise, serology-based antigen-detection can be useful as an approach to diagnose any Leishmania species infection using different corporal fluid samples. Immunogenic and secreted proteins constitute powerful targets for diagnostic methods in antigen detection. As such, we performed immunoproteomic (2-DE, western blot and mass spectrometry) and bioinformatic screening to search for reactive and secreted proteins from L. amazonensis, L. braziliensis, and L. infantum. Twenty-eight non-redundant proteins were identified, among which, six were reactive only in L. amazonensis extracts, 10 in L. braziliensis extracts, and seven in L. infantum extracts. After bioinformatic analysis, seven proteins were predicted to be secreted, two of which were reactive only in L. amazonensis extracts (52kDa PDI and the glucose-regulated protein 78), one in L. braziliensis extracts (pyruvate dehydrogenase E1 beta subunit) and three in L. infantum extracts (two conserved hypothetical proteins and elongation factor 1-beta). We propose that proteins can be suitable targets for diagnostic methods based on antigen detection.

MC3T3-E1 Cell Response to Ti1-xAgx and Ag-TiNx Electrodes Deposited on Piezoelectric Poly(vinylidene fluoride) Substrates for Sensor Applications.

In the sensors field, titanium based coatings are being used for the acquisition/application of electrical signals from/to piezoelectric materials. In this particular case, sensors are used to detect dynamic mechanical loads at early stages after intervention of problems associated with prostheses implantation. The aim of this work is to select an adequate electrode for sensor applications capable, in an initial stage to avoid bone cell adhesion, but at a long stage, permit osteointegration and osteoinduction. This work reports on the evaluation of osteoblast MC3T3-E1 cells behavior in terms of proliferation, adhesion and long-term differentiation of two different systems used as sensor electrodes: Ti1-xAgx and Ag-TiNx deposited by d.c. and pulsed magnetron sputtering at room temperature on poly(vinylidene fluoride) (PVDF). The results indicated an improved effect of Ag-TiNx electrodes compared with Ti1-xAgx and TiN, in terms of diminished cell adhesion and proliferation at an initial cell culture stage. Nevertheless, when cell culture time is longer, cells grown onto Ag-TiNx electrodes are capable to proliferate and also differentiate at proper rates, indicating the suitability of this coating for sensor application in prostheses devices. Thus, the Ag-TiNx system was considered the most promising electrode for tissue engineering applications in the design of sensors for prostheses to detect dynamic mechanical loads.

Alterações histológicas dos rins e expressão das metalotioneínas e das proteínas de choque térmico em ratos Wistar após exposição ao fungicida tirame / Kidney histological alterations and metallothionein and heat shock protein expression in Wistar rats after fungicide thiram exposure

Avaliaram-se as alterações histológicas e a expressão das metalotioneínas (MTs) e das proteínas de choque térmico (Hsp70) nos rins de ratos Wistar após a exposição ao fungicida tirame. Os animais foram distribuídos em três grupos: grupo dieta-padrão; grupo dieta-padrão+óleo de milho; e grupo tirame. Foram encontradas diferenças significativas (P<0,05) na evolução do peso corporal entre os ratos do grupo tirame e os dos grupos controle e óleo de milho, e não foram verificadas lesões histológicas evidentes nos rins dos animais. Foram encontradas diferenças entre os animais do grupo exposto ao tirame e os dos grupos controle e óleo quanto às características histomorfométricas relativas ao corpúsculo renal - exceto para a proporção área da cápsula de Bowman:área do glomérulo - e relativas aos túbulos contorcidos proximal e distal - exceto para altura do epitélio dos túbulos distais. Nos ratos expostos ao tirame, foi observada imunomarcação positiva para as MTs, de moderada a forte, nos túbulos contorcidos da região cortical, diminuindo do córtex em direção à medula, e forte imunomarcação para as Hsp70 nas áreas do córtex e da medula, no glomérulo e nos túbulos contorcidos. Os resultados sugerem que o tirame pode ter toxicidade crônica nos mamíferos por afetar o seu crescimento e que a expressão das MTs e das Hsp70, provável resposta celular adaptativa ao estresse oxidativo causado pelo tirame, pode ser utilizada como biomarcador de exposição a este químico.

The histological alterations and the expression of metallothionein (MTs) and heat shock protein (Hsp70) in the kidney of Wistar rats after thiram fungicide exposure were evaluated. Animals were distributed into three groups: standard diet group, standard diet + corn oil group and thiram group. Significant differences were found (P<0,05) in the evolution of body weight between rats in the thiram group and those in the control and corn oil groups, and no histological lesions were evident in the animals' kidneys. Differences were found among animals in the group exposed to thiram and the control and oil groups regarding histomorphometric characteristics of the renal corpuscle - except for the proportion in the area of Bowman's capsule: glomerulus area - and regarding the height of the epitelium in the distal tubules. In rats exposed to thiram, a positive moderate to strong immunoexpression was observed for MTs, in the cortical convulated tubules decreasing the cortex towards the medulla, and a strong immunoexpression for Hsp70 in the cortex and medulla areas, in the glomerulus and convulated tubules. The results suggest that thiram may have chronic toxicity in mammals affecting their growth, and that the expression of MTs and Hsp70, a probable cellular adaptive response to the oxidative stress caused by thiram, may be used as a biomarker of exposure to this chemical.

Microwave-assisted digestion of organic samples: how simple can it become?

Advancements in sample preparation for performing trace analysis of inorganic analytes are coming from the dissemination of microwave-assisted procedures, but there is still room for improvements by looking for simple and easily applied procedures. Recently it was proposed a new approach called single reaction chamber with capability for digestions at high pressures and temperatures using simple vials and racks. This was a limitation of former cavity microwave ovens with closed vessels. It was demonstrated here that the use of single reaction chamber approach allows the implementation of efficient digestions using diluted solutions of nitric acid and also allows addressing a critical need of sample preparation for inorganic analysis by running mixed batches of samples. The feasibility of this procedure was demonstrated for organic samples and accuracy was proved by using certified reference materials of apple leaves, bovine liver and whole milk powder. Digestions performed of whole milk powder and bovine liver using 2.0 mol L(-1) nitric acid solution plus concentrated hydrogen peroxide at 240 °C led to residual carbon contents of 0.825 and 1.50% and residual acidities of 1.04 and 0.618 mol L(-1), respectively. These parameters are fully compatible with further measurements using ICP OES or ICP-MS. Al, Cu, Fe, Mn, Mo, Rb, Se, Sr, and Zn were accurately determined by ICP OES or ICP-MS depending on their concentrations in digests.

Effect of dithiocarbamate thiram on Wistar rat growth plate and articular cartilage / Efeito do ditiocarbamato tirame sobre a placa de crescimento e cartilagem articular de ratos Wistar

Avaliou-se o efeito do tirame, ditiocarbamato largamente utilizado na agricultura como antifúngico e repelente de roedores, na ossificação endocondral de mamíferos, usando, como modelo, ratos Wistar. Não foram observadas lesões na cartilagem articular, nem nas placas de crescimento, o que pode ser atribuído à dose utilizada e à duração do ensaio. A diminuição da altura da placa de crescimento nos animais aos quais foi administrado o tirame parece traduzir o atraso verificado no crescimento em geral, e não um efeito específico na cartilagem, uma vez que as diferentes zonas da placa epifisária mantiveram as proporções dos animais do grupo-controle. Embora não tenham sido verificados, no presente trabalho, os efeitos registrados para outras espécies nos tecidos cartilaginosos, sugere-se a avaliação dos efeitos crónicos do tirame no crescimento e no desenvolvimento dos ossos longos em mamíferos.

Reduction of heavy metal contents in liquid effluents by vermicomposts and the use of the metal-enriched vermicomposts in lettuce cultivation.

The removal of Cu, Ni and Zn from electroplating effluents by adsorption in cattle manure vermicompost has been discussed. A glass column 38 cm long and 7 cm i.d. was loaded with cattle manure vermicompost and effluents were passed through it. The metal concentrations were measured in the elutant. The experiments on adding effluent aliquots into the columns were continued until the metal concentrations in the elutant reached the maximum values established for effluent discharges in water courses by the Brazilian quality criteria, i.e., Cu=1.0 mg L(-1), Ni=2.0 mg L(-1), and Zn=5.0 mg L(-1). The amount of Cu retention by the vermicompost was determined at the natural effluent pH (2.0). The Zn and Ni retentions were evaluated at the natural effluent pH (6.9 and 7.4, respectively) as well pH 2.0. Vermicompost residues obtained from this process were used for lettuce cultivation. The vermicompost was found to be efficient in removing metals from the electroplating wastes, as well as in the increase of its pH values. Metal retention values were close to 100%. The Cu concentrations in lettuce leaves from the treatment with vermicompost enriched with this metal were below the range of critical toxicity level to plants, i.e., from 20 to 100 mg L(-1). However, the estimated Cu concentrations in the roots from the treatment with vermicompost enriched with Cu were much larger than that of the treatment with the natural vermicompost, reaching 246.3 mg L(-1). The Ni and Zn concentrations in lettuce leaves from the treatments, with vermicomposts enriched with the respective metals, were above the range of critical toxicity levels to plants, i.e., from 10 to 50 mg kg(-1) and from 15 to 30 mg kg(-1), respectively. However, no symptom of toxicity was found visually. Larger accumulations of Cu, Ni and Zn were found in the lettuce leaves than in the roots after the treatments with the uncontaminated vermicompost. A greater absorption of Cu and Ni by roots was found in treatments with vermicompost enriched with these elements, whereas Zn was found preferentially in the leaves. The statistical analysis was done by analyses of variance and regression.

Risk factors for postoperative anxiety in children.

BACKGROUND: Anxiety is defined as a set of behavioural manifestations that can be divided into state- and trait-anxiety. State-anxiety is a transitory emotional condition that varies in intensity and fluctuates over time. Trait-anxiety is a personality trait which remains relatively stable over time. The objective of this study was to identify and quantify perioperative risk factors for immediate postoperative anxiety in children. METHODS: A prospective cohort study was performed with 90 schoolchildren, ages ranging from 7 to 13 years old, ASA physical status I-II, submitted to elective surgery. The measuring instruments were verbal scale of pain, visual analogue scale (VAS), Trait-State Anxiety Inventory for Children (STAIC), Trait-State Anxiety Inventory (STAI) for parents, and structured questionnaire. RESULTS: Patients not submitted to analgesic block and patients with moderate and intense pain presented an estimated risk 5- and 13-fold greater for high levels of postoperative state-anxiety, respectively. High levels of preoperative state-anxiety and administration of doses of midazolam less than 0.056 mg x kg(-1) constituted an estimated risk for postoperative state-anxiety of 3- and 4-fold, respectively. A positive history of previous surgery was associated with lower risk for postoperative anxiety. CONCLUSIONS: High levels of preoperative state-anxiety, administration of less than 0.056 mg x kg(-1) of midazolam, absence of analgesic block and presence of moderate and intense postoperative pain constituted risk factors for immediate postoperative state-anxiety in children. Previous surgery reduced the risk for postoperative anxiety.

A first step incorporating smoking education into a Brazilian medical school curriculum: results of a survey to assess the cigarette smoking knowledge, attitudes, behaviour, and clinical practices of medical students.

Smoking-related illnesses are the leading causes of morbidity and mortality in Brazil. Despite a smoking prevalence of approximately 40%, there is limited national effort to reduce tobacco use in Brazil by means of public education and training of health care professionals to promote smoking education. In particular, the need for information about tobacco warrants increased emphasis in undergraduate medical education. An educational program on nicotine addiction during medical school could facilitate the incorporation of smoking cessation interventions into routine medical practice. As a preliminary step toward implementing a tobacco education and intervention program, this study was designed to assess knowledge and attitudes about smoking among Brazilian medical students. Five hundred thirteen (N = 513) medical students from the Federal University of Rio Grande do Sul, the southernmost state of Brazil, completed a self-reported questionnaire during the 1995-1996 academic school year. Most students recognize the adverse health effects of smoking and the importance of their professional role in promoting smoking cessation. In contradiction, however, few medical students currently provide their patients who smoke with even minimal intervention. This discrepancy supports the idea that training in nicotine addiction and smoking cessation techniques will help medical students to develop the skills and confidence needed to successfully intervene with their current and future patients.

[Drug surveillance: concepts, objectives, and methods]. / Farmacovigilância: conceitos, objectivos e métodos.

Adverse reactions to drugs, by their frequency and morbidity, constitute a very important problem in clinical practice today. However, it is not always possible to establish the definitive causal relationship between the drug and the clinico-laboratory situation. This is not only due to the variability of these findings and the inherent difficulties of the methodology used, but also the fact that most doctors show little sensitivity to such an important subject. In this paper we present a short review of the drug surveillance programme: basic concepts, objectives and the most important methodology, as well as the current situation in the EEC countries.