Phosphoprotein phosphatase 1 (PPP1) is the most widely expressed and abundant serine/threonine protein phosphatase. PPP1 regulates a variety of cellular functions such as glycogen metabolism, mitosis and meiosis, cell-cycle arrest, apoptosis, dynamics of actin cytoskeleton, protein synthesis among others. The versatility of the PPP1 catalytic subunit (PPP1C) is achieved by associating with different regulatory subunits that target PPP1C to a particular subcellular compartment determining its substrate specificity and activity. PPP1C is expressed in mammals in three isoforms: PPP1CA, PPP1CB and PPP1CC, each encoded by distinct genes. The PPP1C gene undergoes alternative splicing to originate PPP1CC1 and PPP1CC2 variants. PPP1A and PPP1C1 are expressed virtually in all tissues but are particularly enriched in the brain. The subcellular localization of the endogenous PPP1C isoforms is not fully elucidated, but all isoforms are found in the nucleus and cytoplasm, despite some isoform-specific differences in intranuclear distribution. Furthermore, it is known that PPP1 plays a key role in mitosis where PPP1C isoforms are differentially targeted to specific subcellular structures. However, in previous studies, non-neuronal cells were used as a model system to study PPP1 distribution during the cell cycle. In the studies here described, we used the neuronal-like cell line SH-SY5Y since PPP1 is a crucial protein in several neuronal functions.

Digitalis purpurea L. and Digitalis thapsi L. (Iberian endemism) are two well known Portuguese species that contain cardiac glycosides. Last year we found some specimens in the Northeast region of Portugal that appeared to us to be natural hybrids between D. purpurea and D. thapsi. In consequence, morphological, genetic and chemical experimental work was performed, involving a minimum of 30 samples of each plant species collected in this geographic area. Preliminary results of this work have been published.

The histological analyzes performed in gonad cells to verify the occurrence of changes in their anatomy is of crucial importance in order to characterize the dynamics of the gonads during the sexual maturation phase of its cells in specimens of wild marine populations. These histological data, together with other reproductive data, can be used in the monitoring of marine environments in which the Solea solea (Ishiba et al., 2008) lives, namely, the coastal waters of the Iberian Peninsula, Africa and Mediterranean sea (Ishiba et al., 2008). Therefore, the main objective of this study was to evaluate the several cellular types and its quantities and anatomies (Ishiba et al., 2008). The study was performed using a total of 19 specimens of the species Solea solea, collected between the months of January and March of 2011 in two different locations of the Portuguese coast (Aveiro and Leixões, Portugal). The gonad samples were fixated in Formalin-acetic acid-alcohol (FAA) and processed in accordance to the standard histology protocol (Culling et al., 1985). The slices of the gonads were colored with Haematoxylin and Eosin (H&E) (used in the study of the general structure of the gonads, as well as in the identification and localization of the different cellular types of the gonad epithelium). As a result of the study, we found 7 different types of gonad cells: I oogonia (It is not photographed ); II a Chromatin nucleolus oocytes (Fig.1. A); II b perinucleolar oocytes (Fig.1. B); III cortical alveoli oocytes (Fig.1. C); IV vitellogenic oocytes (Fig.1. D); V maturing oocyte (Fig.1. E); VI ovulated oocyte (Fig.1. F), (Hibiya, et al., 1995). The different cellular phases were found in all the gonad samples analyzed in this study and the results corresponded to those expected: a decrease in the size in the cells of the first cellular type, the oogonia and an increase in the size in the cells of all other cellular types found in the study. Histological and histochemical analyses of the Solea solea can be used in anatomic and pathological evaluations of the physiological condition of the animals, as stated previously for the muscles, where histological changes could be observed (such as: degeneration and atrophy of the muscle bundles). In the future, the analytical determination of histological and histochemical changes in the gonads of Solea solea, in conjunction with other analytical processes, such as in situ analyzes, can potentially be used to determine and accurately characterize the physiological condition of animals sampled in any natural environment.

Fish renal morphology has been studied since the 1960’s, but is still incomplete for most species, even for some of the most studied. The functional and morphological diversity and specialization of the fish kidney, due to environmental adaptations, makes this a large subject that remains understudied. References to studies on fish renal structure are given in Table 1. Although authors as Hentschel and Elger contributed to the knowledge of fish renal morphology in a comparative perspective, the knowledge concerning this issue is still reduced. Here we give a short comparative summary of fish renal morphology, using as baseline model the brown trout.

The species of earthworms included in the Lumbricidae family have a very complex esophageal organ called calciferous or Morren gland, being the representatives of the genus Lumbricus the individuals where that gland is better developed. Previous studies have shown the secretion produced in the glandular structure is not crystalline but amorphous in the form of amorphous calcium carbonate (ACC) spherulites. Afterwards the spherulites undergo a crystallization process to form rhombohedral calcite crystals that finally result in macroaggregates of these crystals. This crystallization process can go through two intermediate states, vaterite or aragonite. These changes happen on the surface of the amorphous phases.

The physiological condition of Solea solea and Solea senegalensis, caught in the natural environment and in aquaculture fish farms respectively, was determined by analyzing the histological changes occurred in the gills of the animals. These two species were used because both occupy the same ecological niche but only Solea senegalensis is reared in Portuguese aquaculture farms. The analysis was performed using 19 specimens in total from both environments collected between January and March of 2011 in two different locations of the Portuguese coast (Leixões and Aveiro, Portugal). The lesions found in the gills samples corresponded to four types of fusions: basal fusion corresponding to fusions at the base of the secondary lamella; medium fusion corresponding to fusions up to half of the secondary lamella; total fusion corresponding to fusions in the total length of the lamella; apical fusion corresponding to the complete fusion of the entire lamella. Another form of lesion found were lamellar edema which consisted of two different type: apical edema corresponding to a swelling at the apex of the secondary lamella; lamellar edema corresponding to a swelling in the full length of the tissue of the secondary lamella. The last physiological modification found was the alteration of the quantity of mucous cells found and also of mucous released in the primary and secondary lamellae of the gills. The results revealed that the most commonly type of lesions found were: basal fusion (Fig. 1. C); medium fusion (Figure 1 B); apical fusion (Figure 1 A); total fusion (Figure 1 D); lamellar edema (Figure 1 E); apical edema (Figure 1 F) and mucous cells disorganization in the gills (Figure 1 G). These data are very similar to other previously reported results in histopathologic studies, and are most likely to result from: contamination with heavy metals; organic or pesticides contamination; contamination by hydrocarbons (oils, fuels; rapid changes in the temperature of the water; the time of recovery of the fishing nets in the case of Solea solea; contamination of the ice used in the storage process of Solea solea. The histological analyses are very useful to evaluate the physiological condition of the fish. The histological analyses are a very important tool which enables the rapid development of processes and supports sound ecological management decisions in order to protect and/or recover habitats and ecosystems in which these animals live, therefore, improving their physiological condition with obvious benefits as to the human consumer is concerned. The novelty in this study was the use of bottom duelling fish and the comparison established between wild specimens and aquaculture reared specimens. The final conclusion is that several types of contamination occurred both in the natural environment as in the aquaculture fish farms, resulting in the different types of lesions and physiological changes observed during our experiment.

Silver nanoparticles are emerging as a powerful tool in bioimaging applications owing to their unique plasmonic properties i.e., extremely high molar extinction coefficients, resonant Rayleigh scattering and enhanced local electromagnetic fields. Through the optimization of these properties, by controlling composition, size, shape, and interparticle spacing of nanoparticles and their assemblies, highly enhanced local electromagnetic fields in the vicinity of nanoparticles are achievable giving rise to IR, Raman and fluorescence surface enhanced spectroscopies (SEIRS, SERS and MEF, respectively).

There is a growing interest in nanoparticles as carriers of chemotherapeutic agents in order to improve their administration and minimize their side effects. Despite the fact that silver nanoparticles can be conjugated to therapeutic agents, offering additionally advantages due their unique and tunable optical properties, few examples have been described yet.

The Alzheimer’s Amyloid Precursor Protein (APP) is a type 1 transmembranar protein that has been implicated in roles such as cell adherence, survival, migration and differentiation. Although a role in neuritogenesis has been attributed to APP, some contradictory results have been reported regarding the benefits of knocking-down or overexpressing APP. Our preliminary work indicated that pAPP (APP phosphorylated at the S655 residue) may potentially modulate APP-mediated neuronal differentiation, through mechanisms that may involve increased APP trafficking and cleavage, and altered APP-mediated cytoskeleton rearrangements. In order to address the ability of APP and pAPP to mediate neuronal differentiation, we used APP cDNA constructs fused to the Green Fluorescent Protein (GFP): wild-type APP and APP S655 mutants (S655A dephosphomimicking and S655E phosphomimicking mutants). SH-SY5Y neuroblastoma cells, a well documented neuronal-like cell model, were used, and the conditions for their retinoic-acid mediated differentiation and concomitant APP-GFP transfection were first optimized.

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus responsible for painful disabling polyarthritis, undergoing re-emergence in the Indian Ocean and the Asian continent. Travelling and changing patterns of vector distribution, and abundance due to climate changes, make CHIKV a global threat without effective control strategies. One approach to reduce the CHIKV burden is the development of a vaccine. Virus-like particles (VLPs) are a safe and highly effective class of subunit vaccines that mimic the overall structure of virus particles. Accordingly, we aim to produce CHIKV VLPs in a baculovirus-insect cell system, well known for allowing high yields of recombinant protein expression, to be used as a vaccine preparation.

The Anthozoa class is composed solely by marine benthic consumers, some with adaptations that enable them to inhabit even the most extreme habitats, like hydrothermal vents, the polar seas, and even whale falls. Anthozoans, such as corals and anemones, are important modulators of marine habitats in the coral reef communities, being also ecologically relevant due to their abundance and the trophic level they occupy. One of these is Actinia equina, the most conspicuous species in tidal pool communities in European shores, with a broad range of distribution from the North Sea to the Mediterranean and Aegean seas and South Africa. Histological and structural analyses allows for a more detailed biological and ecological study of these organisms, including their reproductive and metabolic behaviour. This work describes a morphologic and histologic analysis of A. equina, with these endpoints for the first time. Organisms, collected to the natural environmental were processed using a paraffin-embedding technique. Further, samples were processed following by Hematoxilin and Eosin (H & E) standard procedures with adaptations to Actiniidae conditions (the fixation time was 96 hours due to 98% of water body in these organisms). Results obtained confirm that, when compared to higher level animals, the Anthozoa are very simply organized (Figure 1). The body is built up of sheets of tissue (Figure 2 and 3), on the surface of each there is an epithelium, resting upon a middle layer of fibrous mesoglea. This generally contains no cellular elements other than some scattered amoebocytes and locally, genital cells (Figure 2). The aim of this work was to contribute to a new anatomic and histological baseline for these sea anemones family Actiniidae. In order to identify several taxonomic characteristics based on reproductive structures. The methods applied were standard to identify several structures that acid-basic groups ligands, such as phosphate of DNA, collagen and other nuclear proteins. The results obtained were the basic cnidarian structures such as tentacular and mesenteric anatomy. The reproductive structures are show in detail with septal filaments and mesenteric (Figure1). These structures is taxonomically relevant and into the own family (Actiniidae) could following the same patterns. The male and female A. equina mesenteries are different. The mainly conclusion of this study was that the male and female gonads can appear simultaneously in the same individual, or that it can have no defined sex.

Since the discovery of the piezoelectric character of bone, the suitability of some piezoelectric materials have been studied for bone repair; they are thought to act like transducers converting the mechanical energy of skeletal deformation in electrical stimuli capable of controlling osteogenic growth. The mechanisms underlying this process are far from being understood and systematic studies at a local scale are required. Atomic force microscopy (AFM) is a unique way to observe phenomena at the nanoscale and liquid imaging provides a unique tool to assess biological phenomena at the nanoscale. So in this study, aiming at a better understanding of the role of piezoelectricity in the osteogenic growth, the interaction between a poled piezoelectric material, in this case poly (L-lactic) acid and an adhesion promoting protein, the fibronectin, and bone-like cells is evaluated by scanning probe microscopy and confocal laser scanning microscopy (CLSM).

Molybdenum disulfide/Tungsten disulphide (MoS2/WS2) is a compound very useful for its properties; it is used as lubricant, catalyst in hydrodesulfuration, in hydrogen fuel storage, etc. As part of the 2nd Joint Congress of the Portuguese and Spanish Microscopy Societies the present work reports about the different types of MoS2/WS2 nanomaterials which have been investigated by using aberration corrected STEM namely: (1) MoS2 nanotubes (2) MoS2 hexagonal nanoplates, (3) rippled or helical MoS2 nanowires, (4) Co-doped MoS2/WS2 nanowires and (5) fullerene-like WS2 nanoparticles.

The effective control over the behavior of nanostructured systems requires a tight control over the shape and size of the nanoparticle building blocks. This control can be very effective and useful in the case of crystalline gold nanoparticles. In order to achieve a precise morphology control over the particles obtained using gold nanorods as seeds; we have studied the crystalline structure of the initial single-crystal gold nanorods, which can be used in subsequent overgrowth processes. We also studied the mechanisms involved in the overgrowth and reshaping of such gold nanorods.

Ever since colossal magnetoresistance (CMR) was discovered in Ln1-xAExMnO3 (Ln=lantanide; AE alkaline-earth), establishing the relationship between structure and properties constitutes a prominent place in the landscape of solid-state science because of its intriguing potential applications. The occurrence of different Mn3+/Mn4+ ratios seems to be responsible of the different magnetic behaviours observed and can be tuned by either changing i) the Ln/AE ratio or ii) the oxygen content, i.e., Ln1-xAExMnO3-δ. Although plenty of works has been devoted to the first point, less effort has been devoted to study the effect of the compositional variations at the oxygen sublattice. In spite of the partial break-up occurring at the 3D network of octahedral sharing corners, responsible of the double exchange interactions the oxygen deficient compounds exhibits complex magnetic and electric properties.

δ-Bi2O3, a material with a fluorite-type structure, is one of the best solid-state oxygen-ion conductors. It is a high-temperature form that cannot be quenched to room temperature. However, doping with small amounts of transition metal oxides preserves the δ-Bi2O3 structure at low temperature and retains its anionic conduction properties. The Bi2O3–MoO3 materials are interesting because of their functional properties, chiefly as catalysts and as good ionic conductors. All the phases in this system are related to the fluorite structure except Bi2MoO6 which shows an Aurivillius-type structure.

A microstructure refined to the nanometer scale originates mechanical property improvements [1]. However ultra-fine grain metals present low thermal stability, requiring the presence of particle dispersions to delay coarsening by grain boundary pinning. Nanoscale dispersions of diamond or graphite offer therefore thermal stability potential, combined with high hardness and thermal conductivity for diamond, and self-lubricating properties in the case of graphite. Copper-diamond (Cu-nD) and copper-graphite (Cu-G), nickel-diamond (Ni-nD) and nickel-graphite (Ni-G), as well as tungsten-diamond (W-nD) and tungsten-graphite (W-G) nanostructured composites have been produced by mechanical synthesis and subsequent heat-treatments. Fundamental challenges involve bonding carbon phases to the chosen matrices: copper exhibits an intrinsically difficult bonding with carbon; while tungsten is a strong carbide former; with Ni exhibiting intermediate characteristics. In addition, carbon phases are prone to amorphization by high-energy milling. Transmission electron microscopy (TEM) observations showed homogeneous particle distributions and intimate bonding between the metallic matrix and the carbon phases. Ring diffraction patterns of the chemically extracted carbon phases demonstrated that nanodiamond milled with Cu preserved crystallinity (Figure 1), while an essentially amorphous nature could be inferred for graphite. Systematic variation of the processing parameters enabled to minimize carbide formation with Ni and W matrices. Heat-treatment of the nanostructured of Ni-nD composites induced the transformation of nanodiamond into onion-like carbons (OLC) inside the Ni nanostructured matrix (Figure 2). Hardening mechanisms and the load transfer ability to the reinforcement particles, as well as thermal stability, have been evaluated through microhardness tests.

The most active phase of the fluid catalytic cracking (FCC) catalyst, used in oil refinery, is zeolite-Y which is an aluminosilicate with a high internal and external surface area responsible for its high reactivity. Waste FCC catalyst is potentially able to be reused in cement-based materials - as an additive - undergoing a pozzolanic reaction with calcium hydroxide (Ca(OH)2) formed during cement hydration. This reaction produces additional strength-providing reaction products i.e., calcium silicate hydrate (C-S-H) and hydrous calcium aluminates (C-A-H) which exact chemical formula and structure are still unknown. Partial replacement of cement by waste FCC catalyst has two key advantages: (1) lowering of cement production with the associated pollution reduction as this industry represents one of the largest sources of man-made CO2 emissions, and (2) improving the mechanical properties and durability of cement-based materials. Despite these advantages, there is a lack of fundamental knowledge on pozzolanic reaction mechanisms as well as spatial distribution of porosity and solid phases interactions at the microstructural level and consequently their relationship with macroscopical engineering properties of catalyst/cement blends.

In recent years there has been an increased interest in characterisation methods capable of evaluating the response of nanostructured materials such as quantum dots and nanotubes to externally applied stimuli. However, due to issues concerning sample homogeneity and non-uniform orientation, the study of collective responses can lead to ambiguous results. Therefore, it is crucial to perform a complementary evaluation at the single nanostructure level which requires the use of sophisticated analytical instrumentation. In addition to high spatial resolution, the equipment employed must permit sample manipulation and real-time imaging of the in situ stimulus response. Transmission electron microscopes (TEM) observe all these requirements. Furthermore, novel developments in sample holder design and manufacturing have opened a window to an unprecedented range of new in situ experiments. As a result, varied reports are increasingly populating the in situ TEM literature examining such different matters as temperature-related structural phase changes of nanoparticles or dopant diffusion in nanoscaled batteries.

Research at the nanometer scale, aiming to create novel materials and devices offering better performance than those of macro/micro systems, is currently of great relevance in many topics of modern science, technology and engineering. In what concerns coatings technology, the nanocomposite films can offer excellent properties in different applications, representing their morphological characterization a great challenge.