Biotechnology Issues Research Articles

Far-Red Fluorescent Proteins: Tools for Advancing In Vivo Imaging.

Far-red fluorescent proteins (FPs) have emerged as indispensable tools in in vivo imaging, playing a pivotal role in elucidating fundamental mechanisms and addressing application issues in biotechnology and biomedical fields. Their ability for deep penetration, coupled with reduced light scattering and absorption, robust resistance to autofluorescence, and diminished phototoxicity, has positioned far-red biosensors at the forefront of non-invasive visualization techniques for observing intracellular activities and intercellular behaviors. In this review, far-red FPs and their applications in living systems are mainly discussed. Firstly, various far-red FPs, characterized by emission peaks spanning from 600 nm to 650 nm, are introduced. This is followed by a detailed presentation of the fundamental principles enabling far-red biosensors to detect biomolecules and environmental changes. Furthermore, the review accentuates the superiority of far-red FPs in multi-color imaging. In addition, significant emphasis is placed on the value of far-red FPs in improving imaging resolution, highlighting their great contribution to the advancement of in vivo imaging.

Developing Supportive Bioeconomy Policies

Developing Supportive Bioeconomy Policies

Designing Active Learning Enriched Biotechnology Instruction Module (BIM) for Middle School Science Course

This study aimed at designing instructional material named Biotechnology Instruction Module (BIM) which is enriched with active learning methods to enhance the 8th grade students’ learning of the topic. The development of the teaching materials proceeded within the stages of the ADDIE instructional design model. The final form of the instructional module comprised active learning materials of DNA Isolation Experiment, Bulletin Boards, 4x2 Biotechnology Learning Station, Case Studies Text, and “What Would You Do If You Were…?” activities. Instructional module covers the topics such as genetically modified organisms (GMO), gene therapy, cloning and bioremediation. The “Biotechnology Instruction Module” is expected to help students to learn in a meaningful way the rapidly changing and renewed biotechnology issues, to foster knowledge and understanding of key issues, to promote socioscientific discourse in class, and to encourage higher order thinking skills. All the data show that the instructional module applied in the real classroom environment is successful in realizing the learning objectives of the subject of biotechnology.

Laser-assisted bioprinting of microorganisms with hydrogel microdroplets: peculiarities of Ascomycota and Basidiomycota yeast transfer.

Laser-assisted bioprinting of microbial cells by hydrogel microdroplets is a rapidly developing and promising field that can contribute to solving a number of issues in microbiology and biotechnology. To date, most research on the use of laser bioprinting for microorganism manipulation and sorting has focused on prokaryotes; the bioprinting of eukaryotic microorganisms is much less understood. The use of hydrogel allows solving two fundamental problems: creating comfortable environments for living microorganisms and imparting the necessary rheological properties of the gel for the stable transfer of microdroplets of a preset size. Two main problems were solved in this article. First, the parameters of the hydrogel based on hyaluronic acid and laser fluence to ensure stable transfer of single drops are selected. Second, possible differences in the bioprinting by hyaluronic acid hydrogel microdroplets with yeasts of various taxonomy (Ascomycota vs Basidiomycota), which form and do not form polysaccharide capsules and evaluated. We have performed laser induced forward transfer of 8 yeast species (Goffeauzyma gilvescens, Lipomyces lipofer, Lipomyces starkey, Pichia manshurica, Saitozyma podzolica, Schwanniomyces occidentalis var. occidentalis, Sterigmatosporidium polymorphum, Vanrija humicola) and assessed its viability based on colony formation on the nutrient medium. It is shown that after laser-induced transfer in hydrogel microdroplets the mean viability rate was 77% with some strains showing relatively high viability rates exceeding 90%. Effect of capsules presence on colony formation after laser bioprinting was not revealed. Differences in laser transfer of the yeast of various phyla were found-basidiomycetes formed a greater number of colonies than ascomycetes. The causes and mechanisms of these effects require detailed studies. The data obtained contributes to the knowledge about the bioprinting of eukaryotic microorganisms and can be useful in the studies of single microbial cells and inter-organism interactions.

생명연장 융합기술에 대한 철학적 성찰

The purpose of this paper is to present a holistic and practicable methodology to address new ethical issues emerging from biotechnology. With the advent of the age of life sciences, related technologies have extended life beyond the purpose of disease treatment without limitation and have even played a role in selecting excellent genes. With the acquisition of this technology by life scientists, mankind has come to encounter ethical problems that are completely different from the past. Ethicists point out that the logic of the age of life sciences has limitations in solving ethical problems. It is pointed out that human beings are in danger of discussing ethical issues that will emerge in the present and future due to bio-science technology. The human community had to call for a new ethical approach. This method is a predictable philosophical reflection on the environment and future of society and the human community beyond the ethical issues of science and technology. Therefore, this research method is not only to approach the ethical issues of biotechnology itself, but also to ensure the possibility of predicting a transparent future

Evaluation of the butyrylcholinesterase expression and activity in CHO, HEK-293 and vero cell lines transformed by dual promoter expression vector

BACKGROUND: Butyrylcholineesterase (BChE) is a therapeutic drug and its producing as a recombinant protein is an essential issue in biotechnology. One of the highlights in this regard is choosing the best host cells and plasmids. OBJECTIVES: The aim of this study is to evaluate the production of butyrylcholinesterase in Vero, HEK-293, and CHO cell lines using a dual promoter vector. MATERIAL AND METHODS: The dual-promoter construction (pBudCE dual BChE) was transfected into cell lines categorized in three experimental groups (pBudCE dual BChE, pCMV and negative control). BChE gene expression and enzyme activity was evaluated at different times. RESULTS: All three cell lines showed higher gene expression level in pBudCE dual BChE group. BChE enzyme activity level of this group in CHO cells decreased in sixth day and increased in ninth day. In HEK-293 cells it has a downward trend from sixth to ninth day and in Vero cells its level in the ninth day was the highest. CONCLUSION: The difference of pBudCE dual BChE and pCMV groups was more pronounced in the HEK-293 cell and the BChE gene expression level of this cells was higher than the others while, CHO cells showed higher level of BChE enzyme activity.

The political economy of China's GMO commercialization dilemma

AbstractWhy has China delayed commercialization of genetically modified (GM) crops over the last decade given its heavy public R&D investment in biotechnology and maturing GM capacity? Further, imports of GM products for consumption are allowed while similar domestic GM‐crops cannot be commercialized. The paper analyzes, from a political economy perspective, how the delay of commercialization arises from the government balancing interests and conflicts among vested interests to keep social stability based on comprehensive secondary data and unique primary survey data. Anti‐GMO activists have, over time, been able to shape the public perception of biotechnology. Their efforts have garnered widespread acceptance among interest groups and have been sufficient to raise the specter of underlying social instability, which is always near the top of mind for China's government. As a result, no movement on commercialization has been made since 2010—more than a decade ago. The failure to commercialize is important because food production would increase considerably if China were able to commercialize GM crops and the resolution of the biotechnology issue has broad implications for food security.

Kinetic Processes in Enzymatic Nanoreactors for In Vivo Detoxification.

Enzymatic nanoreactors are enzyme-encapsulated nanobodies that are capable of performing biosynthetic or catabolic reactions. For this paper, we focused on therapeutic enzyme nanoreactors for the neutralization of toxicants, paying special attention to the inactivation of organophosphorus compounds (OP). Therapeutic enzymes that are capable of detoxifying OPs are known as bioscavengers. The encapsulation of injectable bioscavengers by nanoparticles was first used to prevent fast clearance and the immune response to heterologous enzymes. The aim of enzyme nanoreactors is also to provide a high concentration of the reactive enzyme in stable nanocontainers. Under these conditions, the detoxification reaction takes place inside the compartment, where the enzyme concentration is much higher than in the toxicant diffusing across the nanoreactor membrane. Thus, the determination of the concentration of the encapsulated enzyme is an important issue in nanoreactor biotechnology. The implications of second-order reaction conditions, the nanoreactor’s permeability in terms of substrates, and the reaction products and their possible osmotic, viscosity, and crowding effects are also examined.

Art’s Work in the Age of Biotechnology: Shaping Our Genetic Futures

Art’s Work in the Age of Biotechnology: Shaping Our Genetic Futures

The Effects of Photobioreactor Type on Biomass and Lipid Production of the Green Microalga Monoraphidium pusillum in Laboratory Scale

Mass production of microorganisms, algae among them, for new bioactive compounds and renewable innovative products is a current issue in biotechnology. The greatest challenge of basic research on this topic is to find the best solution for both physiology and scalability. In this study, the main goal was to highlight the contradictions of physiological and technological optimization in the same, relatively small, laboratory scale. The green alga Monoraphidium pusillum (Printz) Komárková-Legnorová was cultured in a conventional Erlenmeyer flask (as air bubbled in a tank-type photobioreactor) and in a hybrid (fermenter type + helical tubular type) photobioreactor of the same volume (2.8 L). Higher cell numbers from 1.7–2.3-fold, 2–2.8-fold higher dry masses, and 1.9–2.6-fold higher total lipid contents (mg·L−1) were measured in the tank reactor than in the hybrid reactor. Cultures in the conventional tank reactor were characterized with better nutrient utilization (42.8–77.7% higher phosphate uptake) and more diverse lipid composition than in the hybrid reactor. The study highlights that well-scalable arrangements and settings could be not optimal (or unsuitable in some cases) from a physiological point of view. The results suggest certain developmental directions for complex, well-scalable devices and highlight the importance of testing the gained physiological optima on these systems.

Ways to reduce the impact of livestock facilities on the environment

The results of the study of the impact of livestock facilities on the state of the environment are presented. To ensure environmental safety, pollutant emissions are regulated by implementing environmental protection policies. Therefore, there is a need to develop scientifically sound approaches to improve methods of regulating emissions of pollutants into the atmosphere from low-capacity livestock facilities, which determines the relevance of this work. The analysis of the existing researches taking into account the European system of inventory of EMEP («CORINAIR») is carried out. A number of measures have been identified to reduce emissions of pollutants into the atmosphere from livestock facilities. It is established that for the last 16 years there is a tendency to reduce the number of farm animals, in particular: cattle by 41.4% and pigs by 21.4%. The exception is poultry, which increased by 28.5% during this period. At the same time, as a result of economic activity of livestock facilities such pollutants as ammonia, hydrogen sulfide, methane, alcohols (methanol, ethanol, etc.), phenols, esters, carbonyl compounds (aldehydes and ketones), carbon acids, sulfides and disulfides, mercaptans, amines, carbon dioxide. The efficiency of introduction of entomological method of utilization of organic waste of animal husbandry which allows receiving simultaneously protein of animal origin and organic fertilizers with the improved physical and mechanical properties is established. Pig manure after processing by larvae of flies becomes a valuable organic fertilizer that has a nematodicidal effect. It is especially valuable for indoor use. Application of biohumus to the soil at the rate of 400 g/m2 reduces the number of nematodes and delays its emergence. Thus, obtaining humus is essentially a solution to the problem of using the ecological mechanism of soil fertility restoration. This addresses the issue of humus biotechnology, which is an alternative to soil chemicalization and creates the conditions for biologization and greening of agriculture. Also, one of the ways to reduce the negative impact on the environment is to dispose of waste in agriculture by producing biogas. This allows you to get a fuel mixture of gases with a heat of combustion of about 20–25 MJ/m3 and methane content in the range of 60–75%, as well as reduce the load of polluting gases on the environment.

Salinity Stress: Toward Sustainable Plant Strategies and Using Plant Growth-Promoting Rhizobacteria Encapsulation for Reducing It

Salinity is one of the most important abiotic stresses that influences plant growth and productivity worldwide. Salinity affects plant growth by ionic toxicity, osmotic stress, hormonal imbalance, nutrient mobilization reduction, and reactive oxygen species (ROS). To survive in saline soils, plants have developed various physiological and biochemical strategies such as ion exchange, activation of antioxidant enzymes, and hormonal stimulation. In addition to plant adaption mechanisms, plant growth-promoting rhizobacteria (PGPR) can enhance salt tolerance in plants via ion homeostasis, production of antioxidants, ACC deaminase, phytohormones, extracellular polymeric substance (EPS), volatile organic compounds, accumulation of osmolytes, activation of plant antioxidative enzymes, and improvement of nutrients uptake. One of the important issues in microbial biotechnology is establishing a link between the beneficial strains screened in the laboratory with industry and the consumer. Therefore, in the development of biocontrol agents, it is necessary to study the optimization of conditions for mass reproduction and the selection of a suitable carrier for their final formulation. Toward sustainable agriculture, the use of appropriate formulations of bacterial agents as high-performance biofertilizers, including microbial biocapsules, is necessary to improve salt tolerance and crop productivity.

Challenges for design of aggregation-resistant variants of granulocyte colony-stimulating factor

Non-native protein aggregation is a long-standing issue in pharmaceutical biotechnology. A rational design approach was used in order to identify variants of recombinant human granulocyte colony-stimulating factor (rhG-CSF) with lower aggregation propensity at solution conditions that are typical of commercial formulation. The approach used aggregation-prone-region (APR) predictors to select single amino acid substitutions that were predicted to decrease intrinsic aggregation propensity (IAP). The results of static light scattering temperature-ramps and chemical unfolding experiments demonstrated that none of the selected variants exhibited improved aggregation resistance, and the apparent conformational stability of each variant was lower than that of WT. Aggregation studies under partly denaturing conditions suggested that the IAP of at least one variant remained unaltered. Overall, this study highlights a general challenge in designing aggregation resistance for proteins, due to the need to accurately predict both APRs and conformational stability.

Recent Developments in Effective Antioxidants: The Structure and Antioxidant Properties.

Since the last few years, the growing interest in the use of natural and synthetic antioxidants as functional food ingredients and dietary supplements, is observed. The imbalance between the number of antioxidants and free radicals is the cause of oxidative damages of proteins, lipids, and DNA. The aim of the study was the review of recent developments in antioxidants. One of the crucial issues in food technology, medicine, and biotechnology is the excess free radicals reduction to obtain healthy food. The major problem is receiving more effective antioxidants. The study aimed to analyze the properties of efficient antioxidants and a better understanding of the molecular mechanism of antioxidant processes. Our researches and sparing literature data prove that the ligand antioxidant properties complexed by selected metals may significantly affect the free radical neutralization. According to our preliminary observation, this efficiency is improved mainly by the metals of high ion potential, e.g., Fe(III), Cr(III), Ln(III), Y(III). The complexes of delocalized electronic charge are better antioxidants. Experimental literature results of antioxidant assays, such as diphenylpicrylhydrazyl (DPPH) and ferric reducing activity power assay (FRAP), were compared to thermodynamic parameters obtained with computational methods. The mechanisms of free radicals creation were described based on the experimental literature data. Changes in HOMO energy distribution in phenolic acids with an increasing number of hydroxyl groups were observed. The antioxidant properties of flavonoids are strongly dependent on the hydroxyl group position and the catechol moiety. The number of methoxy groups in the phenolic acid molecules influences antioxidant activity. The use of synchrotron techniques in the antioxidants electronic structure analysis was proposed.

Industrial Biotechnology is Firing on All Cylinders

Industrial Biotechnology is Firing on All Cylinders

MAINSTREAM NEWSPAPERS COVERAGE OF AGRICULTURAL BIOTECHNOLOGY ISSUES: A STUDY OF DAILY TRUST AND THE NATION NEWSPAPERS

Mainstream newspapers’ coverage of biotechnology issues has overtime attracted the attention of media scholars. This study examined select newspaper’s coverage of Agricultural Biotechnology issues in Nigeria. The study relied on quantitative content analysis of the news stories of 2 Nigerian newspapers (Daily Trust and The Nation newspapers). The study analysed contents of the select newspapers between the periods of September 2018 to March 2019. The study is anchored on the Agenda setting theory. Findings from the study showed that the two newspapers (Daily Trust and The Nation) have not given adequate coverage to the issues of Agricultural Biotechnology as they were episodic in their reportage of the issue. The newspapers were unable to do this because most of the reports on Agricultural Biotechnology are in the form of news writing, paying little attention to the analysis of risks or benefits that lies therein. It was also found that the selected newspapers accorded low prominence to the issues on Agricultural Biotechnology with the placement of a majority of such stories on the inside pages and far less on the front and back pages. In a nutshell, the study found out that the Newspapers studied have been unsuccessful in their role to communicate to the public on the benefits or otherwise of Agricultural Biotechnology. Thus, it was concluded that Daily Trust and The Nation newspapers adopted a similar pattern in the reportage of Agricultural Biotechnology issues during this period with the way they reported these issues in their respective pages. The study recommends that media; both print and broadcast, which have the capacity of shaping public debate and discourse among citizens should be analytic and should dedicate more space and time when reporting core issues on Agricultural Biotechnology in Nigeria.

Saudi students’ and science teachers’ knowledge of and attitudes towards biotechnology

ABSTRACT This study examined Saudi secondary students’ and science teachers’ knowledge of, and attitudes towards, biotechnology. A mixed methodology approach (i.e., incorporating semi-structured interviews and questionnaires) was utilised. A questionnaire was provided to 138 Saudi students (44 grade 10 students, 45 grade 11 students, and 49 grade 12 students) and 75 Saudi science teachers; in addition, a semi-structured interview was carried out with 10 of these science teachers to explore their opinions on biotechnology. The questionnaire was a Likert scale survey comprising 43 questions. Fifteen of these questions (i.e., the Biotechnology Knowledge Questionnaire, BKQ) were linked to biotechnological knowledge as a way to determine the extent of the secondary students’ and science teachers’ knowledge about biotechnology and how it is used; a further 28 questions were added to fulfil the above objective. The findings from this research indicate a limited awareness of biotechnology among Saudi students and science teachers. Furthermore, most of the interviewed educators agree that the Saudi science curriculum insufficiently addresses the most important issues in biotechnology. In order to increase their awareness, students need to be introduced to the subject of biotechnology within an overarching Saudi science education curriculum.

A positive experience in applying the biolistic approach to potato varieties Aksor and Nevskiy.

The method of biological ballistics (biolistic transformation, genetic bombardment) of plants is one of the most modern methods used for direct gene transfer into plant cells. The main advantages of this method include the ability to simultaneously incorporate several target genes into the plant genome, carry out transfer without unnecessary agrobacterial parts and plasmid DNA sequences, and the short time needed to produce transgenic cells. For different plant objects, the efficiency of obtaining transgenic plants by the ballistic method varies from 1 to 3 %. For potato plants, the transformation efficiency is quite low at the moment and the selection of optimal conditions for biolistics is one of the pressing issues of practical biotechnology. This article presents a successful experience of introducing two genes of interest into two potato varieties using the biolistic approach. The results of biolistic transformation experiments are presented for two types of explants: potato internodes and calli of the varieties Aksor and Nevskiy. Of the 862 explants used for transformation, 56 regenerated plants were obtained. PCR screening of transformants revealed one plant with the insertion of the chitinase gene, one with the insertion of the endo-β-1,3-glucanase gene, and co-transformation by both genes was confirmed in four regenerants. The average transformation efficiency for potato explants was 0.7 %. A high number of regenerants (56) as opposed to a low number of transformants (6) reflects an attempt to increase the number of regenerants by using a lower concentration of the selective agent (kanamycin). Although this approach requires more effort, it can be used to produce potato lines with integrated genes of interest for further use in crop breeding. The lines of potato obtained in the current study by introducing two genes associated with the plant response to fungal pathogens will be further assessed for their resistance to fungal diseases and, if successful, will be used in potato crop breeding.

Selected Aspects Of The Crispr-Cas Biology And Applications

Abstract The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins are components of the adaptive immunity system, protecting against foreign DNA, which are present in many bacteria species. Recent years have brought extensive research on this system however, not all of its biological properties have been discovered so far. It was recently discovered that CRISPR-Cas can regulate the formation of biofilm and is closely associated with the DNA repair system in bacterial cells. It is also likely that some of the spacer sequences are complementary to short sequences in the bacterial genome, which may have an influence on regulation of bacterial genes, e.g. virulence factors. Besides, phages can synthesize anti-CRISPR genes, which could be of use in the future for the purpose of development of an alternative therapy against multi-drug resistant bacterial strains. Here we present an elementary characteristic of CRISPR-Cas system, including the structure and the brief mechanism of action, systematic classification and its importance for medicine and biotechnology issues. We would like to stress the huge potential of CRISPR-Cas by discussing the selected but varied aspects. 1. Introduction. 2. Structure, operation and differences. 3. Bacterial typing. 4. Correlation with bacterial pathogenicity. 5. Potential tool for medicine. 5.1. CRISPR-tool for genome editing. 5.2. Instances of CRISPR-tool strategies in medicine. 6. Phage response. 7. Conclusions

Introduction: The Interactive Relations Between Science and Technology and Literary Studies

Introduction: The Interactive Relations Between Science and Technology and Literary Studies