Genome editing technologies and their applications in crop improvement

Crop improvement is very essential to meet the increasing global food demands and enhance food nutrition. Conventional crop-breeding methods have certain limitations such as taking lot of time and resources, and causing biosafety concerns. These limitations could be overcome by the recently emerged-genome editing technologies that can precisely modify DNA sequences at the genomic level using sequence-specific nucleases (SSNs). Among the artificially engineered SSNs, the CRISPR/Cas9 is the most recently developed targeted genome modification system and seems to be more efficient, inexpensive, easy, user-friendly and rapidly adopted genome-editing tool. Large-scale genome editing has not only improved the yield and quality but also has enhanced the disease resistance ability in several model and other major crops. Increasing case studies suggest that genome editing is an efficient, precise and powerful technology that can accelerate basic and applied research towards crop improvement. In this review, we briefly overviewed the structure and mechanism of genome editing tools and then emphatically reviewed the advances in the application of genome editing tools for crop improvement, including the most recent case studies with CRISPR/Cpf1 and base-editing technologies. We have also discussed the future prospects towards the improvement of agronomic traits in crops.     

Bifidobacteria and their role in human health

Summary There is a growing consensus on the beneficial effects of bifidobacteria in human health. It is now clear that bifidobacteria that exist in the large intestine are helpful for maintenance of human health and are far more important thanLactobacillus acidophilus as beneficial intestinal bacteria throughout human life. In other words, the reduction or disappearance of bifidobacteria in the human intestine would indicate an unhealthy state. Oral administration of bifidobacteria may be effective for the improvement of intestinal flora and intestinal environment, for the therapy of enteric and hepatic disorders, for stimulation of the immune response, and possibly for the prevention of cancer and slowing the aging process. However, for consistent and positive results further well-controlled studies are urgently needed.     

Salmonellae in health foods

Various health food products of different brands were purchased from stores in the metropolitan Atlanta area. These foods were examined for the presence of salmonellae by fluorescent-antibody and cultural methods. Included in the study were tablets of alfalfa, parsley, kelp, wheat bran, enzyme, bone meal, and vitamins. Beef liver powder and tablets and granola cereal were also studied. Salmonella minnesota, Salmonella anatum, and Salmonella derby were isolated from two of three lots of beef liver powder from one manufacturer. All other products were negative.     

Applications of cell culture for the improvement of human health

Current applications of Cell Culture Engineering which have a major beneficial impact for the improvement of human health range from a great variety of vaccines (examples include measles, mumps, rubella, polio, and Hepatitis A) made using this technology to a whole new range of therapeutic proteins dependent for their expression on animal cell culture hosts (examples include EPO, TPA and -interferon). Novel applications in the pipeline include cell therapy products and gene therapy products.     

Salmonellae in health foods.

Various health food products of different brands were purchased from stores in the metropolitan Atlanta area. These foods were examined for the presence of salmonellae by fluorescent-antibody and cultural methods. Included in the study were tablets of alfalfa, parsley, kelp, wheat bran, enzyme, bone meal, and vitamins. Beef liver powder and tablets and granola cereal were also studied. Salmonella minnesota, Salmonella anatum, and Salmonella derby were isolated from two of three lots of beef liver powder from one manufacturer. All other products were negative.     

Functional and safety aspects of enterococci in dairy foods

The genus Enterococcus like other LAB has also been featured in dairy industry for decades due to its specific biochemical traits such as lipolysis, proteolysis, and citrate breakdown, hence contributing typical taste and flavor to the dairy foods. Furthermore, the production of bacteriocins by enterococci (enterocins) is well documented. These technological applications have led to propose enterococci as adjunct starters or protective cultures in fermented foods. Moreover, enterococci are nowadays promoted as probiotics, which are claimed for the maintenance of normal intestinal microflora, stimulation of the immune system and improvement of nutritional value of foods. At the same time, enterococci present an emerging pool of opportunistic pathogens for humans as they cause disease, possess agents for antibiotic resistance, and are frequently armed with potential virulence factors. Because of this “dualistic” nature, the use of enterococci remains a debatable issue. However, based on a long history of safe association of particular enterococci with some traditional food fermentations, the use of such strains appears to bear no particular risk for human health. Abundance of knowledge as well as progress in molecular techniques has, however, enabled exact characterization and safety assessment of strains. Therefore, a balanced evaluation of both, beneficial and undesirable nature of enterococci is required. A clear understanding of their status may, therefore, allow their safe use as a starter, or a probiotic strain. The present review describes the broader insight of the benefits and risks of enterococci in dairy foods and their safety assessment.     

Haematococcus astaxanthin: applications for human health and nutrition

The carotenoid pigment astaxanthin has important applications in the nutraceutical, cosmetics, food and feed industries. Haematococcus pluvialis is the richest source of natural astaxanthin and is now cultivated at industrial scale. Astaxanthin is a strong coloring agent and a potent antioxidant - its strong antioxidant activity points to its potential to target several health conditions. This article covers the antioxidant, UV-light protection, anti-inflammatory and other properties of astaxanthin and its possible role in many human health problems. The research reviewed supports the assumption that protecting body tissues from oxidative damage with daily ingestion of natural astaxanthin might be a practical and beneficial strategy in health management.     

Monoclonal antibodies directed against human FcRn and their applications

The MHC class I-like Fc receptor (FcRn) is an intracellular trafficking Fc receptor that is uniquely responsible for the extended serum half-life of antibodies of the IgG subclass and their ability to transport across cellular barriers. By performing these functions, FcRn affects numerous facets of antibody biology and pathobiology. Its critical role in controlling IgG pharmacokinetics has been leveraged for the design of therapeutic antibodies and related biologics. FcRn also traffics serum albumin and is responsible for the enhanced pharmacokinetic properties of albumin-conjugated therapeutics. The understanding of FcRn and its therapeutic applications has been limited by a paucity of reliable serological reagents against human FcRn. Here, we describe the properties of a new panel of highly specific monoclonal antibodies (mAbs) directed against human FcRn with diverse epitope specificities. We show that this antibody panel can be used to study the tissue expression pattern of human FcRn, to selectively block IgG and serum albumin binding to human FcRn in vitro and to inhibit FcRn function in vivo. This mAb panel provides a powerful resource for probing the biology of human FcRn and for the evaluation of therapeutic FcRn blockade strategies.Key words: FcRn, IgG, monoclonal antibody, albumin, therapy     

Functional foods and strategies contrasting bacterial adhesion

Antibacterial strategies targeting bacterial adhesion to substrates are considered a valuable alternative to traditional antibiotic therapy, in view of the great advantage they bring in combating the infectious process at the very early stage without selecting for drug resistant cells. Amongst bioactive compounds with activity against bacterial adhesion, several are found in natural food and beverages, such as cranberry, tea, coffee, wine and milk. For the analysis of their anti-infective potential, successful experimental models can be conducted using different substrates from the oral cavity. Studies conducted so far in this field allowed the discovery of a variety of anti-adhesive fractions and compounds proven to be effective against bacterial traits involved in the development of oral pathologies such as caries and gingivitis/periodontitis. Discovering new anti-adhesive compounds from natural products, unravelling and testing their prophylactic and therapeutic values, and improving their use in the general population are promising new frontiers in the global fight against human infectious diseases.     

Functional variants of the human 5-lipoxygenase gene and their genetic diagnosis

Variants in the 5-lipoxygenase (ALOX5) gene are first-line candidate causes for interindividual differences in diseases where leukotrienes play a key role, e.g., inflammatory and immune diseases, atherosclerosis, asthma or the acute respiratory distress syndrome (ARDS). We developed and validated Pyrosequencing™ screening assays for single nucleotide polymorphism (dbSNP-IDs rs4986832, rs4987105, rs2115819, rs3740107, rs1565096, rs2291427, rs10571382, rs2242334, rs2229136, rs3802548), and a capillary electrophoresis assay for the ALOX5 Sp1/Egr1 promoter tandem repeat polymorphism. This selection spans the whole ALOX5 gene range and includes all variants with reported functional associations. A gene structure analysis in DNAs from 187 healthy unrelated Caucasians revealed two haploblocks, one in the promoter and one spanning six SNPs from rs3740107G>A in intron 6 to rs2229136A>G in exon 13. The five-repeat genotype was the most frequent Sp1/Egr1 promoter tandem repeat variant (allelic frequency 84%). These assays and analyses provide a solid basis for future assessments of the genetic modulation of leukotriene production.     

Functional expression of parasite drug targets and their human orthologs in yeast

Background

The exacting nutritional requirements and complicated life cycles of parasites mean that they are not always amenable to high-throughput drug screening using automated procedures. Therefore, we have engineered the yeast Saccharomyces cerevisiae to act as a surrogate for expressing anti-parasitic targets from a range of biomedically important pathogens, to facilitate the rapid identification of new therapeutic agents.

Methodology/Principal Findings

Using pyrimethamine/dihydrofolate reductase (DHFR) as a model parasite drug/drug target system, we explore the potential of engineered yeast strains (expressing DHFR enzymes from Plasmodium falciparum, P. vivax, Homo sapiens, Schistosoma mansoni, Leishmania major, Trypanosoma brucei and T. cruzi) to exhibit appropriate differential sensitivity to pyrimethamine. Here, we demonstrate that yeast strains (lacking the major drug efflux pump, Pdr5p) expressing yeast (^ScDFR1), human (^HsDHFR), Schistosoma (^SmDHFR), and Trypanosoma (^TbDHFR and ^TcDHFR) DHFRs are insensitive to pyrimethamine treatment, whereas yeast strains producing Plasmodium (^PfDHFR and ^PvDHFR) DHFRs are hypersensitive. Reassuringly, yeast strains expressing field-verified, drug-resistant mutants of P. falciparum DHFR (^Pfdhfr ^51I,59R,108N) are completely insensitive to pyrimethamine, further validating our approach to drug screening. We further show the versatility of the approach by replacing yeast essential genes with other potential drug targets, namely phosphoglycerate kinases (PGKs) and N-myristoyl transferases (NMTs).

Conclusions/Significance

We have generated a number of yeast strains that can be successfully harnessed for the rapid and selective identification of urgently needed anti-parasitic agents.     

Assisted ultrasound applications for the production of safe foods

Ultrasound requires high power and longer treatment times to inactivate micro‐organisms when compared to ultrasound combined with other technologies. Previous reports have shown that the effectiveness of ultrasound as a decontamination technology can be increased by combining it with another treatment such as pressure, heat and antimicrobial solutions. Assisted ultrasound, the combination of ultrasound with another technology, is more energy efficient, and it has less impact on the food properties. In this review paper, the power ultrasound antimicrobial mechanisms of action, the antimicrobial effects of ultrasound in combination with other physical processes and antimicrobial solutions are comprehensively discussed. Furthermore, the present interest on using these technologies as alternative processing and decontamination methods is presented. Research outputs on the application of ultrasound combined with physical processes are showcased including applications of thermosonication, manosonication, manothermosonication and osmosonication. Antimicrobial efficacy, energy requirements and optimal operation conditions of the different assisted ultrasound technologies are critically discussed, and their impact on the food industry for future applications is presented. Overall, this review paper highlights the importance and recent developments of assisted ultrasound for enhancing food safety.