Modulation of the secretome of hBMSCs by tailoring the macromolecular gradient in hydrogels to generate tissue-to-tissue interfaces

Tissue-to-tissue interfaces are commonly present in all tissues exhibiting structural, biological and chemical gradients serving a wide range of physiological functions. These interfaces are responsible for mediation of load transfer between two adjacent tissues. They are also important structures in sustaining the cellular communications to retain tissueâ s functional integration and homeostasis. [1] All cells have the capacity to sense and respond to physical and chemical stimulus and when cultured in three-dimensional (3D) environments they tend to perform their function better than in two-dimensional (2D) environments. Spatial and temporal 3D gradient hydrogels better resemble the natural environment of cells in mimicking their extracellular matrix. [2] In this study we hypothesize that differential functional properties can be engineered by modulation of macromolecule gradients in a cell seeded threedimensional hydrogel system. Specifically, differential paracrine secretory profiles can be engineered using human Bone Marrow Stem Cells (hBMSCâ s). Hence, the specific objectives of this study are to: assemble the macromolecular gradient hydrogels to evaluate the suitablity for hBMSCâ s encapsulation by cellular viability and biofunctionality by assessing the paracrine secretion of hBMSCâ s over time. The gradient hydrogels solutions were prepared by blend of macromolecules in one solution such as hyaluronic (HA) acid and collagen (Col) at different ratios. The gradient hydrogels were fabricated into cylindrical silicon moulds with higher ratio solutions assembled at the bottom of the mould and adding the two solutions consecutively on top of each other. The labelling of the macromolecules was performed to confirm the gradient through fluorescence microscopy. Additionally, AFM was conducted to assess the gradient hydrogels stiffness. Gradient hydrogels characterization was performed by HA and Col degradation assay, degree of crosslinking and stability. hBMSCâ s at P3 were encapsulated into each batch solution at 106 cells/ml solution and gradient hydrogels were produced as previously described. The hBMSCâ s were observed under confocal microscopy to assess viability by Live/Dead® staining. Cellular behaviour concerning proliferation and matrix deposition was also performed. Secretory cytokine measurement for pro-inflammatory and angiogenesis factors was carried out using ELISA. At genomic level, qPCR was carried out. The 3D gradient hydrogels platform made of different macromolecules showed to be a suitable environment for hBMSCâ s. The hBMSCâ s gradient hydrogels supported high cell survival and exhibited biofunctionality. Besides, the 3D gradient hydrogels demonstrated differentially secretion of pro-inflammatory and angiogenic factors by the encapsulated hBMSCâ s. References: 1. Mikos, AG. et al., Engineering complex tissues. Tissue Engineering 12,3307, 2006 2. Phillips, JE. et al., Proc Natl Acad Sci USA, 26:12170-5, 2008

Changes in the floristic composition of a Terra Firme rain forest In Brazilian Amazonia over an eight-year period In response to logging

Changes in the floristic composition over an eight-year period in a logged area at the Tapajós National Forest in Brazilian Amazonia arc discussed. Two treatments of different intensities of logging were compared with an undisturbed (control) forest. Data were collected from permanent sample-plots. The effects of logging on floristic composition were stronger in the more heavily logged treatment. The number of species decreased immediately after logging, but started to increase before the fifth year after logging and was higher at the end of the study period than before logging. The more heavily logged plots responded more to disturbances, as judged by the increase in the number of species during the period after logging. This forest appears to recover its initial floristic composition after disturbance without intervention.

Audit quality: does gender composition of audit firms matter?

Using a sample of Portuguese audit firms and their client companies, this study examines the association between gender composition of the partnership structure oaudit firms and audit quality. Audit quality is measured through the earnings quality of audit clients. We find that gender diversity in the partnership structure of audit firms, per se, has no association with audit quality. In turn, we find significant evidence that a predominant presence of female Certified Public Accountants (CPAs) in partner positions in audit firms is associated with higher audit quality. In particular, the results indicate that audit firms with female-dominated partnership structures are negatively related with aggressive accounting practices in audit clients.

Floristic composition and diversity of Amazonian rainforest bryophytes in the state of Acre, Brazil

Bryophyte diversity in Amazonian Brazil is high, presently recorded at 514 species distributed among 36 families and 89 genera. Comparisions of species richness among the Amazonian states of Brazil suggests that the bryoflora of Acre is still underestimated, presently recorded at 106 species distributed among 33 families and 63 genera. After two field trips, the diversity increased 50% showing that the bryoflora is still poorly known. Collection data and comments on morphology, ecology and distribution are given for sixty-two species of bryophytes collected in the state of Acre, Brazil, which are new (fifty-two species) or interesting floristic records (ten species). Microlejeunea crenulifolia (Gottsche) Steph. and Stictolejeunea balfourdii Mitt. are new records for Brazil and fifty species are new records for Acre.

Structure and composition of the ground-herb community in a terra-firme Central Amazonian forest

The herb community of tropical forests is very little known, with few studies addressing its structure quantitatively. Even with this scarce body of information, it is clear that the ground herbs are a rich group, comprising 14 to 40% of the species found in total species counts in tropical forests. The present study had the objective of increasing the knowledge about the structure and composition of the ground-herb community and to compare the sites for which there are similar studies. The study was conducted in a tropical non-inundated and evergreen forest 90 km north of Manaus, AM. Ground herbs were surveyed in 22 transects of 40 m², distributed in five plots of 4 ha. The inventoried community was composed of 35 species, distributed in 24 genera and 18 families. Angiosperms were represented by 8 families and Pteridophytes by 10 families. Marantaceae (12 sp) and Cyperaceae (4 sp) were the richest families. Marantaceae and Poaceae were the families with greatest abundance and cover. Marantaceae, Poaceae, Heliconiaceae and Pteridophytes summed 96% of total herb cover, and therefore were responsible for almost all the cover of the community. The 10 most important species had 83.7% of the individuals. In general, the most abundant species were also the most frequent. Richness per transect varied from 7 to 19 species, and abundance varied from 30 to 114 individuals. The community structure was quite similar to 3 other sites in South America and one site in Asia.

Associations between energetic expenditure, physical activity and body composition: exploratory study with industrial workers

This study, which involved a target population comprised by 292 workers of different industrial areas (metalmechanics, foundry, chemical, wood, food), aimed to verify the association between energy expenditure-EE, physical activity level-PAL and body composition (Body Mass Index-BMI, Waist-Hip Ratio-WHR and Waist To Height Ratio, WTHR) of participants. The work was completed with the description of the variables relating to the gender of the individuals (male and female) and the activities carried out in the two sectors of industrial work (administrative sector and productive sector). In this research, the statistical technique of principal components analysis (PCA) and the hierarchical analysis of clusters (HCA) were used. Sociodemographic and anthropometric data were collected as well as the level of physical activity and energy expenditure were assessed. The vast majority of individuals who spend greater energy expenditure and has more intense physical activity were male. Most of these workers are in the production sector. We can confirm that that both, gender and labor activity, are factors that have influence on the EE and the PAL.

Essential oil composition of leaf and fine stem of Aniba canelilla (Kunth) Mez from Manaus, Brazil

The essential oil of the leaves and fine stems of Aniba canelilla (Kunth) Mez collected in the city of Manaus, AM, Brazil, were obtained by hydrodistillation and analyzed by GC/MS. Forty-two components were identified, of which 1-nitro-2-phenylethane, as expected, was the major (71.2%-68.2%).

Chemical composition and acaricidal activity of the leaf and fruit essential oils of Protium heptaphyllum (Aubl.) Marchand (Burseraceae)

Essential oils from leaves and fruits of Protiumheptaphyllum collected in Tamandaré beach Pernambuco/Brazil were analysed by GC/MS and tested for toxicity and repellent effect against the two spotted spider mite (Tetranychus urticae). The major constituent identified in the fruits was alpha-terpinene (47.6 %) whereas oil from leaf contained mainly sesquiterpenes such as 9-epi-caryophyllene (21.4 %), trans-isolongifolanone (10.7 %) and 14-hydroxi-9-epi-caryophyllene (16.7 %). The fruit oil was found to be more effective against the mite when compared to the leaf oil. Both showed mortality properties and oviposition deterrence in higher concentration (10 µl.l-1 air), but only the essential oil from fruits induced repellence on T. urticae.

Influence of glucose concentration on the structure and quantity of biofilms formed by Candida parapsilosis

Candida parapsilosis is nowadays an emerging opportunistic pathogen and its increasing incidence is part related to the capacity to produce biofilm. In addition, one of the most important C. parapsilosis pathogenic risk factors includes the organisms\textquoteright selective growth capabilities in hyper alimentation solutions. Thus, in this study, we investigated the role of glucose in C. parapsilosis biofilm modulation, by studying biofilm formation, matrix composition and structure. Moreover, the expression of biofilm-related genes (BCR1, FKS1 and OLE1) were analyzed in the presence of different glucose percentages. The results demonstrated the importance of glucose in the modulation of C. parapsilosis biofilm. The concentration of glucose had direct implications on the C. parapsilosis transition of yeast cells to pseudohyphae. Additionally, it was demonstrated that biofilm related genes BCR1, FKS1 and OLE1 are involved in biofilm modulation by glucose. The mechanism by which glucose enhances biofilm formation is not fully understood, however with this study we were able to demonstrate that C. parapsilosis respond to stress conditions caused by elevated levels of glucose by up-regulating genes related to biofilm formation (BCR1, FKS1 and OLE1).

The effects of solvent composition on the affinity of a peptide towards hair keratin: experimental and molecular dynamics data

The study of the interaction between hair filaments and formulations or peptides is of utmost importance in fields like cosmetic research. Keratin intermediate filaments structure is not fully described, limiting the molecular dynamics (MD) studies in this field although its high potential to improve the area. We developed a computational model of a truncated protofibril, simulated its behavior in alcoholic based formulations and with one peptide. The simulations showed a strong interaction between the benzyl alcohol molecules of the formulations and the model, leading to the disorganization of the keratin chains, which regress with the removal of the alcohol molecules. This behavior can explain the increase of peptide uptake in hair shafts evidenced in fluorescence microscopy pictures. The model developed is valid to computationally reproduce the interaction between hair and alcoholic formulations and provide a robust base for new MD studies about hair properties. It is shown that the MD simulations can improve hair cosmetic research, improving the uptake of a compound of interest.

Chemical composition of the essential oils from two subspecies of Protium heptaphyllum

Qualitative and quantitative analyses of the volatile constituents from resin of Protium heptaphyllum (Aubl.) Marchand subsp. ulei (Swat) Daly (PHU), and Protium heptaphyllum (Aubl.) Marchand subsp. heptaphyllum (PHH), Burseraceae were performed using GC-MS and GC-FID. The resins were collected around the city of Cruzeiro do Sul, state of Acre, Brazil. Essential oils from the two subspecies were extracted by hydrodistillation with a yield of 8.6% (PHU) and 11.3% (PHH); the main components were terpinolene (42.31%) and p-cymene (39.93%) for subspecies ulei (PHU) and heptaphyllum (PHH), respectively.

Antioxidant potential of two Apiaceae plant extracts: a comparative study focused on the phenolic composition

The present study aimed to characterize the extracts prepared from Pimpinella anisum L. (anise) and Coriandrum sativum L. (coriander) (Apiaceae plants) seeds in terms of phenolic composition, and to correlate the obtained profiles with the antioxidant activity. Anise gave the highest abundance in phenolic compounds (42.09± 0.11 mg/g extract), mainly flavonoids (28.08±0.17 mg/g extract) and phenolic acids (14.01±0.06 mg/g extract), and also the highest antioxidant potential, accessed for the ability to inhibit lipid peroxidation and -carotene bleaching, reducing power and free radical scavenger activity. Apigenin and luteolin derivatives, as also caffeoylquinic acid derivatives appear to be directly related with the higher in vitro antioxidant potential of the anise extract.. In contrast, the weak antioxidant potential of coriander seems to be due to their lower abundance in phenolic compounds (2.24±0.01 mg/g extract). Further studies are necessary to evaluate the in vivo antioxidant potential of the tested extracts, but the performed in vitro experiments highlight them as potential health promoters.

Diversity of sand flies in domiciliary environment of Santarém, state of Pará, Brazil: species composition and abundance patterns in rural and urban areas

This paper aimed to evaluate the richness, abundance and frequency of sand fly occurrence in rural and urban areas American visceral Leishmaniasis -AVL is endemic in the study area of Santarém municipality, Pará state. Sand flies were collected during 1995-2000, using CDC light traps placed in neighborhoods and rural areas of the municipality. A total of 53.454 individuals and 26 species of sand flies were collected. The most abundant species in both urban and rural environments was Lutzomyia longipalpis, vector of AVL in the area. The highest species richness by capture was in rural area. In all years sampled, the largest number of species of sand fly collected was always in rural areas. The species of sand flies in urban and rural area were similar in 11 species. In the rural area other 11 species were found, a total of 22 species. Shannon-Wiener index ranged from 0.12 to 0.84 at rural areas and 0.08 to 0.34 at urban ones. In general, rural localities showed higher diversity (H') of phlebotomines than urban ones. Individual-based rarefaction curves for each area demonstrated that urban localities had the lowest expected number of phlebotomine species and the richest rural ones reach higher expected values with lower amount of individuals than urban sites. The most frequent species were Lutzomyia longipalpis, Evandromyia carmelinoi and Bichromomyia flaviscutellata.

Ichthyometry and electrical bioimpedance analysis to estimate the body composition of tambatinga

Body composition analysis is relevant to characterize the nutritional requirements and finishing phase of fish. The aim of this study was to investigate the relationship between ichthyometric (weight, total and standard length, density and yields), bromatological (fat, protein, ash and water content) and bioelectrical-impedance-analysis (BIA) (resistance, reactance, phase angle and composition indexes) variables in the hybrid tambatinga (Colossoma macropomum × Piaractus brachypomus). In a non-fertilized vivarium, 520 juveniles were housed and fed commercial rations. Then, 136 days after hatching (DAH), 15 fish with an average weight of 37.69 g and average total length of 12.96 cm were randomly chosen, anesthetized (eugenol) and subjected to the first of fourteen fortnightly assessments (BIA and biometry). After euthanasia, the following parts were weighed: whole carcass with the head, fillet, and skin (WC); fillet with skin (FS); and the remainder of the carcass with the head (CH). Together, FS and CH were ground and homogenized for the bromatological analyses. Estimates of the body composition and yields of tambatinga, with models including ichthyometric and BIA variables, showed correlation coefficients ranging from 0.81 (for the FS yield) to 1,00 (for the total ash). Similarly, models that included only BIA variables had correlation coefficients ranging from 0.81 (FS and CH yields) to 0.98 (for the total ash). Therefore, in tambatinga, the BIA technique allows the estimation of the yield of the fillet with skin and the body composition (water content, fat, ash, and protein). The best models combine ichthyometric and BIA variables.

Chemical composition of phytoplankton from the estuaries of Eastern Amazonia

Phytoplankton is important bioindicator of chemical and biological modifications of natural ecosystems. The objective of this study was to determine the total chemical composition of the phytoplankton of the Pará and Mocajuba estuaries on the eastern coast of the Amazon region in the Brazilian state of Pará. The chemical composition of the surface water, bottom sediments (total sample and bioavailable fraction), and the phytoplankton were determined by inductively coupled plasma optical emission spectrometry. Phytoplankton contained high concentrations of Ca, P, Mn, Fe, Zn, Al, Ba, and Pb. The phytoplankton of the Mocajuba estuary is rich in Fe (2,967-84,750 µg g-1), while those from the Pará is rich in Al (1,216-15,389 µgg-1), probably reflecting divergent anthropogenic inputs. Both samples indicated a high bioconcentration factor derived from both the water and the bioavailable fraction, reflecting the efficiency of these organisms in the concentration of metals.