Production of first generation adenoviral vectors for preclinical protocols: amplification, purification and functional titration

Gene therapy represents a promising approach in the treatment of several diseases. Currently, the ideal vector has yet to be designed; though, adenoviral vectors (Ad-v) have provided the most utilized tool for gene transfer due principally to their simple production, among other specific characteristics. Ad-v viability represents a critical variable that may be affected by storage or shipping conditions and therefore it is advisable to be assessed previously to protocol performance. The present work is unique in this matter, as the complete detailed process to obtain Ad-v of preclinical grade is explained. Amplification in permissive HEK-293 cells, purification in CsCl gradients in a period of 10 h, spectrophotometric titration of viral particles (VP) and titration of infectious units (IU), yielding batches of AdβGal, AdGFP, AdHuPA and AdMMP8, of approximately 1013-101? VP and 1012-1013 IU were carried out. In vivo functionality of therapeutic AdHuPA and AdMMP8 was evidenced in rats presenting CCl?-induced fibrosis, as more than 60% of fibrosis was eliminated in livers after systemic delivery through iliac vein in comparison with irrelevant AdβGal. Time required to accomplish the whole Ad-v production steps, including IU titration was 20 to 30 days. We conclude that production of Ad-v following standard operating procedures assuring vector functionality and the possibility to effectively evaluate experimental gene therapy results, leaving aside the use of high-cost commercial kits or sophisticated instrumentation, can be performed in a conventional laboratory of cell culture.     

Thermal wet oxidation improves anaerobic biodegradability of raw and digested biowaste

Anaerobic digestion of solid biowaste generally results in relatively low methane yields of 50-60% of the theoretical maximum. Increased methane recovery from organic waste would lead to reduced handling of digested solids, lower methane emissions to the environment, and higher green energy profits. The objective of this research was to enhance the anaerobic biodegradability and methane yields from different biowastes (food waste, yard waste, and digested biowaste already treated in a full-scale biogas plant (DRANCO, Belgium)) by assessing thermal wet oxidation. The biodegradability of the waste was evaluated by using biochemical methane potential assays and continuous 3-L methane reactors. Wet oxidation temperature and oxygen pressure (T, 185-220 degrees C; O2 pressure, 0-12 bar; t, 15 min) were varied for their effect on total methane yield and digestion kinetics of digested biowaste. Measured methane yields for raw yard waste, wet oxidized yard waste, raw food waste, and wet oxidized food waste were 345, 685, 536, and 571 mL of CH/g of volatile suspended solids, respectively. Higher oxygen pressure during wet oxidation of digested biowaste considerably increased the total methane yield and digestion kinetics and permitted lignin utilization during a subsequent second digestion. The increase of the specific methane yield for the full-scale biogas plant by applying thermal wet oxidation was 35-40%, showing that there is still a considerable amount of methane that can be harvested from anaerobic digested biowaste.     

A Capillary Electrophoresis Method for the Determination of Hydroxyproline as a Collagen Content Index in Meat Products

The addition of collagen or its hydrolysates as a protein source or water binding agent is a common practice in the production of meat products. However, its level of addition is restricted by international regulations. Glycine, proline, and hydroxyproline are the most abundant amino acids in collagen, which is the main constituent of connective tissue. Thus, the objective of this study was to develop a method for the determination of hydroxyproline content in meat products by capillary electrophoresis (CE) as a collagen content index. Acid hydrolysis of samples and amino acid derivatization with fluorescamine prior to CE analysis were conducted. Separations were carried out using an uncoated capillary at 33 °C, using tetraborate buffer 0.05 M, pH 9.3 with 100 mM SDS. Hydroxyproline was detected at 214 nm. The assay was reproducible (RSD < 3% for normalized peak areas and migration times) and had good correlation (r = 0.975, p < 0.001) with the AOAC official method. From the samples tested (36), commercial ham showed the lowest average collagen content (<7.16 g per 100 g protein) by the CE method. On the other hand, frankfurter and Mexican “chorizo” sausages showed average collagen contents of 12.88 and 11.17 g per 100 g protein, respectively. The CE method developed could be used by regulatory agencies to ensure compliance with maximum levels of collagen addition in processed meats.     

Dysfunctional Gut Microbiome Networks in Childhood IgE-Mediated Food Allergy

The development of food allergy has been reported to be related with the changes in the gut microbiome, however the specific microbe associated with the pathogenesis of food allergy remains elusive. This study aimed to comprehensively characterize the gut microbiome and identify individual or group gut microbes relating to food-allergy using 16S rRNA gene sequencing with network analysis. Faecal samples were collected from children with IgE-mediated food allergies (n = 33) and without food allergy (n = 27). Gut microbiome was profiled by 16S rRNA gene sequencing. OTUs obtained from 16S rRNA gene sequencing were then used to construct a co-abundance network using Weighted Gene Co-expression Network Analysis (WGCNA) and mapped onto Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. We identified a co-abundance network module to be positively correlated with IgE-mediated food allergy and this module was characterized by a hub taxon, namely Ruminococcaceae UCG-002 (phylum Firmicutes). Functional pathway analysis of all the gut microbiome showed enrichment of methane metabolism and glycerolipid metabolism in the gut microbiome of food-allergic children and enrichment of ubiquinone and other terpenoid-quinone biosynthesis in the gut microbiome of non-food allergic children. We concluded that Ruminococcaceae UCG-002 may play determinant roles in gut microbial community structure and function leading to the development of IgE-mediated food allergy.     

Optimization of microwave pretreatment on wheat straw for ethanol production

An orthogonal design (L₉(3⁴)) was used to optimize the microwave pretreatment on wheat straw for ethanol production. The orthogonal analysis was done based on the results obtained from the nine pretreatments. The effect of four factors including the ratio of biomass to NaOH solution, pretreatment time, microwave power, and the concentration of NaOH solution with three different levels on the chemical composition, cellulose/hemicellulose recoveries and ethanol concentration was investigated. According to the orthogonal analysis, pretreatment with the ratio of biomass to liquid at 80 g kg⁻¹, the NaOH concentration of 10 kg m⁻³, the microwave power of 1000 W for 15 min was confirmed to be the optimal condition. The ethanol yield was 148.93 g kg⁻¹ wheat straw at this condition, much higher than that from the untreated material which was only 26.78 g kg⁻¹.     

Comparison of environmental impacts of building structures with in situ cast floors and with precast concrete floors

In this paper, the environmental impacts of two types of slab systems are studied. The first type, a concrete-based one-way spanning slab, is the most common solution in residential buildings in Spain. The second type, a hollow core slab floor, is increasingly becoming more used in buildings, but is not being broadly used in residential buildings as yet. The study is undertaken through the Life Cycle Analysis methodology, applying the EPS 2000 method. The assessment shows that the environmental impact of a building structure with precast concrete floors is 12.2% lower than that with in situ cast floors for the defined functional unit.     

Safety of milk‐derived bioactive peptides

Bioactive milk peptides are protein fragments with potential biological activities. Over the past decades, studies have mainly focused on the health benefits of these bioactive peptides; however, only a few studies have evaluated their safety. In this review, we describe the scientific literature regarding their effects on both cell lines and on animal and human models, in terms of allergenicity, cytotoxicity, genotoxicity and mutagenicity.     

Chemical compounds and mechanisms involved in the formation and stabilization of foam in sparkling wines

The visual properties of sparkling wine including foam and bubbles are an indicator of sparkling wine quality. Foam properties, particularly foam height (FH) and foam stability (TS), are significantly influenced by the chemical composition of the wine. This review investigates our current knowledge of specific chemical compounds and, the mechanisms by which they influence the foam properties of sparkling wines. Grape and yeast proteins, amino acids, polysaccharides, phenolic compounds, organic acids, fatty acids, ethanol and sugar are examined with respect to their contribution to foam characteristics in sparkling wines made with the Traditional, Transfer, and Charmat and carbonation methods. Contradictory results have been identified that appear to be due to the analytical methods used to measure and quantify compounds and foam. Biopolymer complexes are discussed and absent knowledge with regards to thaumatin-like proteins (TLPs), polysaccharides, amino acids, oak-derived phenolic compounds and organic acids are identified. Future research is also likely to concentrate on visual analysis of sparkling wines by in-depth imaging analysis and specific sensory analysis techniques.     

Opportunity or Loss? Health Impacts of Estate Renewal and the Relocation of Public Housing Residents

Estate renewal programs can have positive and negative impacts on the health and wellbeing of public housing residents. We aimed to investigate the potential health impacts of relocating residents as part of the redevelopment of a public housing estate in South Western Sydney, New South Wales. In-depth interviews were undertaken with public housing residents as well as with informants from local health and housing authorities. We found that a range of current and potential health impacts were linked to individual residents’ responses to the processes of estate redevelopment and relocation, housing and neighbourhood quality, residents’ social networks and access to social and health care services. Improved housing quality and maintenance, a personalised approach to relocation and equitable access to services were identified to contribute to positive health outcomes.