We present a theoretical model and experimental demonstration of thin liquid film deformations due to a dielectric force distribution established by surface electrodes. We model the spatial electric field produced by a pair of parallel electrodes and use it to evaluate the stress on the liquid–air interface through Maxwell stresses. By coupling this force with the Young–Laplace equation, we obtain the deformation of the interface. To validate our theory, we design an experimental set-up which uses microfabricated electrodes to achieve spatial dielectrophoretic actuation of a thin liquid film, while providing measurements of microscale deformations through digital holographic microscopy. We characterize the deformation as a function of the electrode-pair geometry and film thickness, showing very good agreement with the model. Based on the insights from the characterization of the system, we pattern conductive lines of electrode pairs on the surface of a microfluidic chamber and demonstrate the ability to produce complex two-dimensional deformations. The films can remain in liquid form and be dynamically modulated between different configurations or polymerized to create solid structures with high surface quality.

We experimentally demonstrate quadrupolar electro-osmotic flows around charged dielectric microspheres immersed in an electrolyte when subjected to an alternating current electric field. We present an electrokinetic model that predicts the flow characteristics based on the phenomena of surface conductance and polarization of the electrolyte concentration around the particles. We refer to these flows as concentration polarization electro-osmosis. We anticipate that these flows may play a major role in the electric-field-induced assembly of colloids and on the electrokinetic manipulation of dielectric micro- and nanoparticles.

Housekeeping genes (HKG) are paramount for accurate gene expression analysis during preimplantation development. Markedly, quantitative reverse transcription polymerase chain reaction (RT-qPCR) in ovine embryos currently lacks HKGs. Therefore, we tested 11 HKGs for RT-qPCR normalization during ovine parthenogenetic preimplantation development. Seven HKGs reached the qPCR efficiency threshold (97.20–105.96%), with correlation coefficients ranging from −0.922 to −0.998 and slopes from −3.22 to −3.59. GeNorm ranked glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and TATA-binding protein (TBP) as the best HKG pair, while H3 histone, family 3A (H3F3A) was the third HKG. Relative gene expression was measured for zinc finger protein X-linked (ZFX) and developmental pluripotency-associated 3 (DPPA3) transcripts during ovine parthenogenetic preimplantation development. ZFX did not show any transcript abundance fluctuation among oocytes, cleavage-stage embryos, and morulae. DPPA3 transcript abundance was also similar among all developmental stages, therefore suggesting that it may not display a maternal gene expression profile. In silico analysis of ovine DPPA3 mRNA and protein showed high conservation to bovine orthologues. However, DPPA3 orthologues differed in regulatory motifs. In conclusion, GAPDH, TBP and H3F3A are stable HKGs in ovine parthenogenetic embryos and allow accurate RT-qPCR-based gene expression analysis.

Sensitive and selective detection for cancer biomarkers is critical in cancer clinical diagnostics. In this work, we report an electrochemical detection platform for the carbohydrate antigen tumor marker 15-3 (CA15-3). It is based on a composite material of poly [2-methoxy-5- (2-ethylhexyloxy) -1,4-phenylenevinylene] (MEH-PPV) and multi-walled carbon nanotubes (MWCNT), which deposited as active layer of an organic thin-film transistor. This layer was surface functionalized with the Anti-CA15-3 monoclonal antibody. The devices have a favorable electrical output response for VSD source-drain potentials between 0 to 5 volts, and VG as 8 volts. Once the antigen (CA15-3) is recognized by the antibody, the electrical response is diminished. The test has a linear response in the concentration range of 0–30 U mL - 1 of CA 15–3, with a lower detection limit of ~1 U mL - 1 and a stability of 90% with respect to the initial values after storing the device for two weeks. The method was successfully applied to the determination of CA15-3 in serum samples. Possibly, this used composite material has a greater scope and can be applied to another type of detection scheme.

The aim of this work was to use X-ray diffraction to identify substances used for adulteration of raw milk and to determine if crystallographic analysis can detect extraneous substances in milk. Two unknown substances were sent anonymously by employers linked to the dairy chain, who claimed that they were added directly in milk prior to water addition by truck drivers. The samples were analyzed by X-ray diffraction and submitted to physicochemical analysis. The first substance was identified by X-ray diffraction as sodium citrate, complying with its physicochemical attributes, such as the powerful ability to decrease the freezing point. The second substance was identified by X-ray diffraction as sucrose and this result was also in agreement with its ability to increase the density, decrease the freezing point and finally, to be positive for sucrose in the resorcinol qualitative test. To evaluate if X-ray diffraction can detect extraneous substances already mixed in milk, fresh raw milk samples tampered with urea, sodium hydroxide, sodium citrate and sucrose were freeze dried and analyzed by X-ray diffraction, with no detection of any extraneous substances at any percentage. This is the first report of attempted diagnosis of extraneous substances in milk by X-ray diffraction. However, this technique can be useful only when applied to identify substances used for adulteration prior to its dilution in milk, since the amorphous nature of milk seems to be a limitation for the accurate detection of extraneous substances.

In the current study, phage-exposed mimotopes as targets against tegumentary leishmaniasis (TL) were selected by means of bio-panning cycles employing sera of TL patients and healthy subjects, besides the immune stimulation of peripheral blood mononuclear cells (PBMCs) collected from untreated and treated TL patients and healthy subjects. The clones were evaluated regarding their specific interferon-γ (IFN-γ) and interleukin-4 (IL-4) production in the in vitro cultures, and selectivity and specificity values were calculated, and those presenting the best results were selected for the in vivo experiments. Two clones, namely A4 and A8, were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of IFN-γ, IL-2, IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. In conclusion, we presented a strategy to identify new mimotopes able to induce Th1 response in PBMCs from TL patients and healthy subjects, and that were successfully used to protect against L. amazonensis infection.

Leishmaniases is a tropical disease caused by protozoa of the genus Leishmania for which the current treatment is expensive, besides increasing reports of parasite resistance. This study investigated the anti-Leishmania amazonensis activity of the essential oil from Aloysia gratissima (AgEO) and guaiol, the major sesquiterpene constituent in the oil. Our results showed that AgEO killed promastigotes and intracellular amastigotes at an IC50 of 25 and 0·16 µg mL−1, respectively, while guaiol killed amastigotes at an IC50 of 0·01 µg mL−1. Both AgEO and guaiol were safe for macrophages up to 100 µg mL−1, as evaluated by the dehydrogenase activity, membrane integrity and phagocytic capacity. AgEO and guaiol did not induce nitrite oxide (NO) in resting macrophages and inhibited the production of NO in lipopolysaccharide-stimulated macrophages. The ultrastructural analysis suggested that AgEO and guaiol act directly on parasites, affecting promastigotes kinetoplast, mitochondrial matrix and plasma membrane. Together, these results pointed out that AgEO and guaiol could be promising candidates to develop anti-Leishmania drugs.

This is the first report on the presence of acanthocephalan parasite Echinorhynchus truttae in brown trout (Salmo trutta) from Spain. A total of 343 fish were captured by local anglers from 19 rivers in the adjacent Tambre and Ulla watersheds in Galicia (north-western Spain). Macroscopic and microscopic analyses of the intestinal contents revealed the presence of adults and/or eggs of E. truttae in 53 of the 123 trout from Tambre river basin (43.1%). By contrast, parasitic forms of this acanthocephalan were only observed in eight of the 220 fish from Ulla basin (3.6%), showing significant differences between the prevalences obtained in two adjacent watersheds (P < 0.001, odds ratio 19.0). Prevalence was significantly higher in specimens >3 years (length >26.0 cm) than in younger specimens (P < 0.05). The absence of Gammarus pulex in the region suggests that native gammarid species in Galicia (Echinogammarus lusitanicus and Echinogammarus beriyoni) may act as intermediate host in the life cycle of E. truttae. Different prevalences of E. truttae indicate that the presence/abundance of the intermediate crustacean host may be different in the two river basins, probably as a consequence of various abiotic factors and anthropogenic activities.

The quality of dietary lipids in the maternal diet can programme the offspring to diseases in later life. We investigated whether the maternal intake of palm oil or interesterified fat, substitutes for trans-unsaturated fatty acids (FA), induces metabolic changes in the adult offspring. During pregnancy and lactation, C57BL/6 female mice received normolipidic diets containing partially hydrogenated vegetable fat rich in trans-unsaturated fatty acids (TG), palm oil (PG), interesterified fat (IG) or soyabean oil (CG). After weaning, male offspring from all groups received the control diet until day 110. Plasma glucose and TAG and liver FA profiles were ascertained. Liver mitochondrial function was accessed with high-resolution respirometry by measuring VO2, fluorimetry for detection of hydrogen peroxide (H2O2) production and mitochondrial Ca2+ uptake. The results showed that the IG offspring presented a 20 % increase in plasma glucose and both the IG and TG offspring presented a 2- and 1·9-fold increase in TAG, respectively, when compared with CG offspring. Liver MUFA and PUFA contents decreased in the TG and IG offspring when compared with CG offspring. Liver MUFA content also decreased in the PG offspring. These modifications in FA composition possibly affected liver mitochondrial function, as respiration was impaired in the TG offspring and H2O2 production was higher in the IG offspring. In addition, mitochondrial Ca2+ retention capacity was reduced by approximately 40 and 55 % in the TG and IG offspring, respectively. In conclusion, maternal consumption of trans-unsaturated and interesterified fat affected offspring health by compromising mitochondrial bioenergetics and lipid metabolism in the liver.

Currently, little is known about the helminth fauna in sirenian species. Therefore, the objective of this study was to assess the frequency of infection by Pulmonicola cochleotrema in Antillean manatees (Trichechus manatus manatus), in the North-eastern region of Brazil. Between the years of 1989 and 2014, 88 manatees found on the North-eastern Brazilian coast were clinically examined. They included animals that were found dead, animals maintained in captivity and specimens reintroduced into conservation areas. During their physical examination, helminths present in necropsied carcasses and in reintroduced animals were collected, as well as faecal samples. Parasites were detected in 7.95% (7/88) of the animals; all specimens collected being identified as P. cochleotrema. Only adult manatees were infected, and in two cases clinical signs were observed. This is the first report on the occurrence of P. cochleotrema in Antillean manatees in the states of Paraíba and Sergipe, in the North-eastern coast of Brazil.

Ecologists examine diet composition in order to assess the spatial and temporal variations in interactions between species, the impact of different species traits on the ecological network structure, and the long-term effects of the removal of different species by small-scale fisheries. In this study, our goal was to compare the diets of silky sharks (Carcharhinus falciformis) and scalloped hammerhead sharks (Sphyrna lewini) off the south-west coast of Mexico in order to infer their diet preferences and spatial distributions. We sampled 164 S. lewini (96 stomachs had food, 68 were empty) and 183 C. falciformis (30 stomachs had food, 153 were empty) in Puerto Madero, Chiapas in 2011. The large number of empty stomachs may be the result of using longline fishing gear, which causes high stress resulting in regurgitation. Based on the index of relative importance (%IRI), the fish Chloroscombrus orqueta (IIR = 27.7%) was the most important species in the diet of S. lewini, while the squid Dosidicus gigas (IIR = 34%) was the primary prey of C. falciformis. Levin's index (Bi) and Shannon's index (H′) confirm that both sharks are generalists, as in other regions. The trophic levels of S. lewini (TL = 4.1) and C. falciformis (TL = 4.2) are characteristic of tertiary consumers; meanwhile, the Morisita–Horn index indicates low interspecific overlap between all categories. These results confirm that these two sharks have different foraging preferences or movement patterns; thus, there is no trophic overlap between species as they play unique roles in the ecological network off the south-west coast of Mexico.

The degree of development and operability of the indicators for the Marine Strategy Framework Directive (MSFD) using Descriptor 1 (D1) Biological Diversity was assessed. To this end, an overview of the relevance and degree of operability of the underlying parameters across 20 European countries was compiled by analysing national directives, legislation, regulations, and publicly available reports. Marked differences were found between countries in the degree of ecological relevance as well as in the degree of implementation and operability of the parameters chosen to indicate biological diversity. The best scoring EU countries were France, Germany, Greece and Spain, while the worst scoring countries were Italy and Slovenia. No country achieved maximum scores for the implementation of MSFD D1. The non-EU countries Norway and Turkey score as highly as the top-scoring EU countries. On the positive side, the chosen parameters for D1 indicators were generally identified as being an ecologically relevant reflection of Biological Diversity. On the negative side however, less than half of the chosen parameters are currently operational. It appears that at a pan-European level, no consistent and harmonized approach currently exists for the description and assessment of marine biological diversity. The implementation of the MSFD Descriptor 1 for Europe as a whole can therefore at best be marked as moderately successful.

Antibodies (Ab) recognizing G-protein coupled receptors, such as β1 and β2 adrenergic (anti-β1-AR and anti-β2-AR, respectively) and muscarinic cholinergic receptors (anti-M2-CR) may contribute to cardiac damage, however their role in chronic chagasic cardiomyopathy is still controversial. We describe that Trypanosoma cruzi-infected C3H/He mice show increased P and QRS wave duration, and PR and QTc intervals, while the most significant ECG alterations in C57BL/6 are prolonged P wave and PR interval. Echocardiogram analyses show right ventricle dilation in infected animals of both mouse lineages. Analyses of heart rate variability (HRV) in chronically infected C3H/He mice show no alteration of the evaluated parameters, while C57BL/6 infected mice display significantly lower values of HRV components, suggesting autonomic dysfunction. The time-course analysis of anti-β1-AR, anti-β2-AR and anti-M2-CR Ab titres in C3H/He infected mice indicate that anti-β1-AR Ab are detected only in the chronic phase, while anti-β2-AR and anti-M2-CR are observed in the acute phase, diminish at 60 dpi and increase again in the chronic phase. Chronically infected C57BL/6 mice presented a significant increase in only anti-M2-CR Ab titres. Furthermore, anti-β1-AR, anti-β2-AR and anti-M2-CR, exhibit significantly higher prevalence in chronically T. cruzi-infected C3H/He mice when compared with C57BL/6. These observations suggest that T. cruzi infection leads to host-specific cardiac electric alterations.