Mineralization of chitosan membrane using a double diffusion system for bone related applications

Chitosan membranes were subjected to a pre-treatment in a double diffusion system, with a calcium solution in one chamber and a phosphate solution in the other chamber. Both chambers were separated by the chitosan membrane and subject to three mineralization periods (5, 10 and 15 minutes). After this pre-treatment the bioactivity of the different calcium phosphate coatings formed was tested for different periods of immersion time, 7, 14 and 21 days at room temperature and 37ºC, in acellular simulated body fluid (1.0x). The results obtained demonstrated that the calcium phosphate coatings formed during the pre-treatment process are bioactive. It was found that the calcification is effective just in the side of the membrane exposed to the calcium solution chamber. This enabled to develop membranes with asymmetric osteoinductive properties that can be useful in different orthopedic applications

A two-stage process for conversion of brewer’s spent grain into volatile fatty acids through acidogenic fermentation

This work is focused on the valorization of brewer’s spent grains (BSG) into volatile fatty acids (VFA) through acidogenic fermentation. VFAs are building blocks for several applications, such as bioplastics’ production. Using acid hydrolysis as pre-treatment, several batch assays were performed and the impact of organic load (OL) and pH on VFA production from BSG hydrolysate was assessed. Regardless of the condition, the produced acids were mainly butyric and acetic acids followed by propionic acid. The OL had a direct impact on the total organic acid concentration with higher concentrations at the highest OL (40 gCOD L-1). pH affected the concentration of individual organic acid, with the highest fermentation products (FP) diversity attained at pH 5.0 and OL of 40 gCOD L-1. To assess the potential application of organic acids for biopolymers (such as polyhydroxyalkanoates) production, the content in hydroxybutyrate (HB) and hydroxyvalerate (HV) monomers was estimated from the respective precursors produced at each pH and OL. The content in HV precursors increased with pH, with a maximum at pH 6.0 (ca. 16% C-mol basis). The acidogenic fermentation of BSG hydrolysate was also assessed in continuous operation, using an expanded granular sludge bed reactor (EGSB). It was shown that the BSG hydrolysate was successfully converted to VFAs without pH control, achieving higher productivities than in the batch operation mode.publishersversionpublishe

Brewers' spent grain as substrates for production of cellulolytic and hemicellulolytic enzymes by different Aspergillus species

Brewers' spent grain (BSG) is the major by-product of the brewing industry, representing around 85% of the total by-products generated. It is a lignocellulosic material containing about 38% cellulose, 29% hemicellulose, chiefly arabinoxylans, and 13% lignin. The production of cellulolytic and hemicellulolytic enzymes using this material as substrate represent an eco-friendly strategy for the lignocellulosic biomass hydrolysis, generating fermentable sugars that can be converted into high- added value products, such as bioethanol, lactic acid, xylitol and others. Thus, this work aimed to evaluate the potential of cellulolytic and hemicellulolytic enzymes production by some Aspergillus species cultivated in BSG. Fungi were grown in minimum media, pH 6.5, with 1% BSG and inoculum was done with 107spores/mL, cultivated at 30°C, 120 rpm, for 5 days. Every 24 hours 2 mL of the samples were collected. The enzymatic activity was performed after the incubation of the crude extract with 1% Linear arabinan, Xylan from beechwood, Xyloglucan, Locust bean gum and CMC, at 50°C for 60 minutes and the reducing sugars were determined using dinitrosalicylic acid (DNS). Synthetic substrates (2 mM of PNP--L-arabinofuranoside, PNP--D-xylopyranoside, PNP--Dglucopyranoside and PNP--D-cellobioside) were also used at the same conditions. The extract from A. niveus showed the best arabinanase (0.284 U/mL) and -glucosidase (0.126 U/mL) activities after 48 and 96 hours of cultivation, respectively. On the other hand, the extract from A. brasiliensis presented the best activities of -L-arabinofuranosidase (0.129 U/mL), -xylosidase (0.265 U/mL) and xylanase (2.15 U/mL) when cultivated for 48 hours. After 72 hours, this fungus also showed the best activities for xyloglucanase (1.06 U/mL), mannanase (0.617 U/mL) and endoglucanase (0.254 U/mL). The extract produced by A. flavus presented the best cellobiohydrolase activity with 0.113 U/mL after 120 hours of cultivation. It is important to mention that A. awamori, A. clavatus and A. terreus also showed good levels of different enzymes produced but they were not the best producers. These data suggest the great potential of different cellulolytic and hemicellulolytic enzymes production using BSG as substrate, which represents an eco-friendly destination for the residues and can generate high-added value products with great biotechnological application.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institutos Nacionais de Ciência e Tecnologia (INCT)Fundação para a Ciência e a Tecnologia (FCT)info:eu-repo/semantics/publishedVersio

Production of an enzymatic cocktail by Aspergillus awamori grown on corn straw with stirred tank bioreactor

Increased agroindustrial activity has led to the accumulation of large amounts of lignocellulosic material (LCM). LCM is nature's most abundant source of renewable carbon, representing a valuable industrial substrate with potential for many applications. Thus, the objective of this work was to screen for different holocellulases and to analyze the production of an Aspergillus awamori enzyme cocktail grown in corn straw (CS) using a stirred tank bioreactor. Screening was performed with A. clavatus, A. flavus, A. terreus, A. niveus, A. awamori and A. brasiliensis cultivated in minimal medium (MM), with 1% CS at 30 ºC or 37 ºC (for A. niveus), 120 rpm, for 5 days. Xylanase (XYN) and endoglucanase (EG) activities were evaluated by formation of reducing sugars using dinitrosalicylic acid (DNS). Cellobiohydrolase (CBH), -glucosidase (BGL) and -xylosidase (BXL) were determined by cleavage of PNP--D-cellobioside, PNP--D-glucopyranoside and PNP--D- xylopyranoside, respectively. After screening, a pre-inoculum was prepared with the best enzyme producer using a 500 mL MM flask and incubated at 30 °C, 120 rpm for 48 hours. The increase of enzyme production was performed in a Benchtop BioFlo 310 bioreactor, with 4.5 L of MM and 1% of CS, and was then inoculated the best enzyme producer. Cultivation was performed at 30 °C, pH 6.5, 275 rpm, air flow 2 v.v.m., for 5 days. During the screening, all fungi presented EG, CBH, BGL, XYN and BXL activities. However, A. awamori was chosen to continue the experiments because of its BXL activity which was 12.6 times higher than that produced by A. niveus. At the scale-up stage, XYN production (47.80 U/ mL) increased 4.1-fold compared to flask activity (11.52 U/mL). BXL also showed 1.6-times higher activity, as well as EG, CBH and BGL, which improved 2.3, 3.3 and 1.2 times their activities, respectively. It was concluded that the staggering of cocktail production improved the enzymatic activities and that corn straw is an excellent source of induction. Furthermore, this cocktail has the potential to be applied in the hydrolysis of different LCM due to the range of holocellulases present.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institutos Nacionais de Ciência e Tecnologia (INCT)Fundação para a Ciência e a Tecnologia (FCT)info:eu-repo/semantics/publishedVersio

Family caregivers: what do they need? An integrative review

Abstract OBJECTIVE Aimed to identify the main needs expressed by family caregivers in caring for adependent person. METHOD An integrative review of the literature in the period between 2010 and 2015 using specific search engine tools in the EBSCO and SCOPUSdatabases. RESULTS 11 articles were selected, and the analysis of the scientific evidence obtained allowed for organizing the results into five thematic areas:transition into care, being responsible for everything, the importance of support, access to formal support, communication and informationprocesses. CONCLUSION The results showed that caregivers have many needs in different areas, which should be addressed in nursing interventions

Evidence for Reductive Genome Evolution and Lateral Acquisition of Virulence Functions in Two Corynebacterium pseudotuberculosis Strains

Ruiz JC, D'Afonseca V, Silva A, et al. Evidence for Reductive Genome Evolution and Lateral Acquisition of Virulence Functions in Two Corynebacterium pseudotuberculosis Strains. PLoS ONE. 2011;6(4): e18551.Background: Corynebacterium pseudotuberculosis, a Gram-positive, facultative intracellular pathogen, is the etiologic agent of the disease known as caseous lymphadenitis (CL). CL mainly affects small ruminants, such as goats and sheep; it also causes infections in humans, though rarely. This species is distributed worldwide, but it has the most serious economic impact in Oceania, Africa and South America. Although C. pseudotuberculosis causes major health and productivity problems for livestock, little is known about the molecular basis of its pathogenicity. Methodology and Findings: We characterized two C. pseudotuberculosis genomes (Cp1002, isolated from goats; and CpC231, isolated from sheep). Analysis of the predicted genomes showed high similarity in genomic architecture, gene content and genetic order. When C. pseudotuberculosis was compared with other Corynebacterium species, it became evident that this pathogenic species has lost numerous genes, resulting in one of the smallest genomes in the genus. Other differences that could be part of the adaptation to pathogenicity include a lower GC content, of about 52%, and a reduced gene repertoire. The C. pseudotuberculosis genome also includes seven putative pathogenicity islands, which contain several classical virulence factors, including genes for fimbrial subunits, adhesion factors, iron uptake and secreted toxins. Additionally, all of the virulence factors in the islands have characteristics that indicate horizontal transfer. Conclusions: These particular genome characteristics of C. pseudotuberculosis, as well as its acquired virulence factors in pathogenicity islands, provide evidence of its lifestyle and of the pathogenicity pathways used by this pathogen in the infection process. All genomes cited in this study are available in the NCBI Genbank database (http://www.ncbi.nlm.nih.gov/genbank/) under accession numbers CP001809 and CP001829

Expression of APOBEC3G/3F and G-to-A Hypermutation Levels in HIV-1-Infected Children with Different Profiles of Disease Progression

OBJECTIVE: Increasing evidence has accumulated showing the role of APOBEC3G (A3G) and 3F (A3F) in the control of HIV-1 replication and disease progression in humans. However, very few studies have been conducted in HIV-infected children. Here, we analyzed the levels of A3G and A3F expression and induced G-to-A hypermutation in a group of children with distinct profiles of disease progression. METHODOLOGY/PRINCIPAL FINDINGS: Perinatally HIV-infected children were classified as progressors or long-term non-progressors according to criteria based on HIV viral load and CD4 T-cell counts over time. A group of uninfected control children were also enrolled in the study. PBMC proviral DNA was assessed for G-to-A hypermutation, whereas A3G and A3F mRNA were isolated and quantified through TaqMan® real-time PCR. No correlation was observed between disease progression and A3G/A3F expression or hypermutation levels. Although all children analyzed showed higher expression levels of A3G compared to A3F (an average fold of 5 times), a surprisingly high A3F-related hypermutation rate was evidenced in the cohort, irrespective of the child's disease progression profile. CONCLUSION: Our results contribute to the current controversy as to whether HIV disease progression is related to A3G/A3F enzymatic activity. To our knowledge, this is the first study analyzing A3G/F expression in HIV-infected children, and it may pave the way to a better understanding of the host factors governing HIV disease in the pediatric setting