Biotechnology Center Research Articles

Establishment and development of the Center of Plant Systems Biology and Biotechnology in Plovdiv, Bulgaria.

The Bulgarian research landscape, presented mainly by the research institutes that are part of the Bulgarian Academy of Sciences and the Agricultural Academy, needs diversification to match the research and innovation potential of the other European Union (EU) countries. This article describes the establishment of the Center of Plant Systems Biology and Biotechnology (CPSBB), a new innovative type of independent research organization that is changing the research landscape in Bulgaria. Supported by the EU Commission, Bulgarian Government, and Plovdiv Municipality, CPSBB has quickly become the leading plant science institute in Bulgaria, creating knowledge in diverse fields such as bioinformatics, biotechnology, genetics and genomics, metabolomics, and systems biology. We outline the organizational structure of CPSBB, the development of its infrastructure, and its scientific productivity. Finally, we compare CPSBB with other similar research establishments in Europe and we conclude that such new types of institutes have a bright future in Bulgaria due to their operational flexibility, productivity, and connections with academia and industry.

Serological Monitoring of Pandemic Influenza Virus Markers in the Russian Federation in 2021–2023

State Scientific Center of Virology and Biotechnology “Vector” has been monitoring highly pathogenic influenza since 2005.The aim of this work was to track the markers of highly pathogenic influenza in the blood sera of people who had a contact with infected and/or deceased birds, as well as of residents from regions where emergence of new variants of influenza A virus is most likely to occur.Materials and methods. Sera were studied using hemagglutination inhibition test (HI test). HI-positive sera were subjected to virus neutralization reaction.Results and discussion. In 2021, 2076 blood serum samples from 19 regions of Russia were collected. Only 7 samples demonstrated significant titers in HI test with A/H5N8 viruses. In 2022, 1620 blood serum samples from 23 regions were obtained; 25 of them were positive for influenza А/H5N8 and А/H5N1 viruses. In 2023 (January-August), 3335 serum samples from 31 regions of the Russian Federation were collected. 28 samples were positive for influenza А/H5N8 and А/H5N1 viruses. Furthermore, we monitored blood sera for low-pathogenic A/H9N2 virus. The number of positive samples in 2021 was lower than 1 % (13 out of 2076); in 2022, it reached 5 % (81 out of 1620); in 2023, the share was lower than 1 % (31 out of 3335). The data obtained suggest indirectly that currently there is no stable circulation of zoonotic influenza A/H5N8 and A/H5N1 viruses in Russia. Influenza viruses A/H9N2 have widely spread in many countries of the world and actively participate in evolution of highly pathogenic influenza A/H5Nx viruses. The Russian Federation demonstrates a gradual increase in the number of blood serum samples with antibodies to A/H9N2 virus.

Key challenges in the dietary intake structure and cutting edge technologies for optimizing nutrition to protect the health of the Russian рopulation

This article presents an analysis of some of the results of the work of the Federal Research Center for Nutrition and Biotechnology (Center) in recent years, highlighting the most important, promising areas of Nutrition Science and Food Hygiene that need further development. The priority area of Center functioning is scientific support for the implementation of the Doctrine of Food Security of the Russian Federation (Decree of the President of the Russian Federation dated January 21, 2020 No. 20), Decree of the President of the Russian Federation dated July 21, 2020 No. 474 «On the national development goals of the Russian Federation for the period until 2030 «in terms of ensuring an increase in life expectancy and improving the life quality of the population, the Strategy for Improving the Quality of Food Products in the Russian Federation until 2030 (Order of the Government of the Russian Federation dated June 29, 2016 No. 1364-r). The Center coordinates all research on medical nutrition problems in the Russian Federation within the framework of the work of the Problem Commission on Nutrition Hygiene of the Scientific Council of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare, the Scientific Council of the Russian Academy of Sciences on Medical Nutrition Problems, the Scientific and Technical Committee of the Comprehensive Scientific Program «Priority Research in the Field of Nutrition of the Population», Profile Commission on Dietetics of the Expert Council in the Field of Health of the Ministry of Healthcare of Russian Federation, ensuring the implementation of their results with the participation of members of the Consortium "Healthcare, Nutrition, Demography". The most important area of the Center's work is scientific and expert support in the field of international and national technical regulation of the production and turnover of foods and raw materials, in particular, the work of the Russian national contact point of the Codex Alimentarius Commission (established by FAO and WHO), as well as the work of the Russian side in the Eurasian Economic Commission regarding the preparation of proposals for technical regulations of the Customs Union in the field of food safety, evaluation of draft technical regulations and amendments and additions to them.

Yield Attributes and Biochemical Compositions of Elites and Landraces of Pigeon Pea [Cajanus cajan (L.) Millsp

Pigeon pea (Cajanus cajan L. Millsp.) is an important grain legume grown mostly in the tropical regions of the world, Nigeria inclusive. The crop is still underutilised in Nigeria; the potentials of the crop in terms of yield traits and biochemical compositions, with ability of enhancing food security and preventing over-dependent on very expensive animal protein, is yet to be fully exploited. It was based on this premise that this study was designed. A total of twenty-five accessions were obtained from International Institute of Tropical Agriculture, lbadan; National Centre for Genetic Resources and Biotechnology, Ibadan and Local farmers in Oyo state, Nigeria. The seeds of the accession were sown in the field in a Randomised Complete Block Design (RCBD) with three replicates. Standard procedures were followed for determination of yield parameters as well as biochemical compositions. It showed that NG/SA/11/08/108 produced the highest grain yield (437.00). Similarly, TCC-8126 produced the highest crude protein (22.53 %) while PG/OY/18/002 had the highest carbohydrate contents (52.22 %). TCC-8126 was the best elite genotype in terms of amino acid profile, containing glutamic acid (18.10 %), aspartic acid (11.4 %); arginine (7.41 %), alanine (5.90 %). The values produced for both nutritional compositions and amino acid profiles were within the recommended standard limits set by Food and Agriculture Organisation and United States Department of Agriculture. NG/SA/11/08/108, NG/AO/MAY/09/021/02, TCC-151, TCC-8126, TT-8129 and TCC-2 have been selected as promising genotypes that could serve as breeding lines for future breeding programme of Cajanus cajan in Nigeria.

Gamma Irradiation Doses reduce Cooking time and Improve Nutritional Profile of selected Pigeon pea [Cajanus cajan (L.) Millsp] Genotypes in Nigeria

Pigeon Pea (Cajanus cajan L.) is a self-pollinating perennial legume shrub with over 30,000 accessions deposited world-wide. Despite the nutritional importance of pigeon peas, its detrimental traits such as its long cooking time, has led to low cultivation of the crop and inhibit its consumers’ request. Concerted efforts to improve both the quality and quantity of this crop through mutation breeding informed this research. It was based on this premise that the study was aimed to evaluate gamma irradiation influences on the nutrient quality and cooking duration of selected pigeon pea varieties in Nigeria. Four accessions of pigeon peas (‘NG/SA/11/08/108’, ‘NG/AO/MAY/00/021/01’, ‘TCC-2’ and ‘NG/SA/07/191’) were obtained from National Centre for Genetic Resources and Biotechnology (NACGRAB), Ibadan. They were irradiated at the Oncological Department ABU Teaching Hospital, Shika, Zaria, using Cobalt-60 (60CO) irradiation source. Standard procedures were followed to determine the effects of the irradiation on cookability and nutritional profile of the M1 plants and their respective controls. It was established that all the parameters tested were irradiation dose dependent. In NG/AO/MAY/00/021/01, 100 Gy produced the highest protein (20.40 %) and highest carbohydrate (68.93 %) contents. Anti-nutritional factors such as cyanide were lower (48.83, 46.22, 43.65 and 44.75 mg/kg in NG/AO/MAY/00/021/01, NG/SA/11/08/108, NG/SA/07/191 and TCC-2) at 400 Gy than those of their respective controls (54.83, 56.83, 53. and 53.55 mg/kg). The amount of Magnesium increased with higher doses of gamma irradiation (1.37 g/100 g for 200 Gy NG/AO/MAY/00/021/01; 1.03 g/100g for 400 Gy NG/SA/11/08/108 and 1.13 g/100 g for TCC-2). There were significant reductions in cooking time in 400 Gy treatments for all the genotypes. The differences between the control and the least cooking time in 400 Gy were 11, 16, 22 and 15 minutes for NG/AO/MAY/00/021/01, NG/SA/11/08/108, NG/SA/07/191 and TCC-2, respectively. It was therefore concluded that these variations could be exploited for further improvement of the crop for food security. Keywords: Cooking time, Mutation breeding, Gamma irradiation, Cookability, Dose dependent

ADVANCING THE PRODUCTION OF VIRAL VECTORS FOR CAR-T CELL THERAPY IN KAZAKHSTAN

Therapy of hematological tumors with chimeric antigen receptor-expressing T-cells (CAR-T) is a recent technology which holds promise to become the most significant achievement in oncohematology over the past 20 years. One aspect of the clinically applied CAR-T technology is that the process of production of CAR+ cells requires viral vectors which are used to deliver the CAR gene in T lymphocytes. At present the production of CAR-T therapy depends on the availability of packaged lentiviral or retroviral vectors. There is a worldwide deficiency in a production capacity to produce CAR vectors, and clinical-use vectors are not sold as off-the-shelf products. At the National center for biotechnology the CAR receptor was assembled and this CAR targets the CD19 antigen which is a marker of tumor cells of the B-cell origin, lentiviral vectors were constructed. Studies on the production of packaged vectors were conducted at which three types of transfection reagents were compared for efficiency, in terms of the produced functional titers. A method was developed to measure functional titers using flow cytometry. High titers of the packaged vectors were obtained. As the CAR-T is effective for treatment of patients after failure of traditional therapy, with relapse or refractory disease, all works on transfer of the CAR-T technology to Kazakhstan are of life importance to the patients with blood cancers in the country. The results underscore a necessity to organize a full production process to produce CAR vectors for clinical use and therapeutic cells for the CAR-T therapy at the NCB.

Identificación de fuentes de resistencia a <i>Fusarium oxysporum</i> en genotipos de <i>Physalis </i>sp.

The resistance of Cape gooseberry to Fusarium oxysporum was evaluated in 70 accessions of Physalis peruviana and related taxa (P. philadelphica, P. ixocarpa, P. floridana, P. pubescens, P. angulata, P. pruinosa, P. viscosa, P. mexicana, Nicandra physalodes, and Solanum auriculatum). These accessions were obtained from different national and international collections, including accessions from the Colombian productive sector. These represented wild materials, commercial cultivars, native and foreign weeds, and commercial ecotypes from the main producing countries: Colombia, South Africa, Kenya, and Peru. The evaluation of resistance to F. oxysporum was carried out under greenhouse conditions using the most aggressive strain supplied by the Fusarium collection maintained by the molecular microbiology laboratory of the Center for Biotechnology and Bioindustry (CBB), currently in charge of the Working Collection of Microorganisms of Agrosavia, which was isolated from infected fields. The symptoms were monitored using a severity scale, containing 10 degrees and five categories. Data information obtained from daily evaluations was analyzed through a severity evaluation and different statistical analyses. The results identified one accession belonging to Physalis peruviana and two related taxa (Physalis floridana and Solanum auriculatum) as resistant to this pathogen. These accessions could be directly used in breeding programs, either as improved cultivars or as race-specific resistance donors for other Physalis peruviana genotypes.

Evaluation of humoral immune responses of experimental animals to the recombinant SARS-CoV-2 spike ectodomain with the ISCOM adjuvant

Scientific relevance. The use of recombinant antigens in vaccine production is limited because vaccines based on such antigens tend to have low immunogenicity. However, a COVID-19 vaccine that combines recombinant SARS-CoV-2 spike glycoprotein as its antigen and virus-like immune-stimulating complexes (ISCOMs) as its adjuvant (Nuvaxovid) induces a protective virus-neutralising response. The State Research Center of Virology and Biotechnology “Vector” (hereinafter, Vector) has developed the ISCOM adjuvant Matrix-V, which plays a key role in inducing virus-neutralising antibodies. Studying Matrix-V will provide for the wide use of recombinant antigens combined with this adjuvant in the development and production of novel Russian vaccines.Aim. This study aimed to evaluate the humoral immune responses of experimental animals to intramuscular injections of a complex combining the recombinant Wuhan-type SARS-CoV-2 spike RBD antigen and the virus-like ISCOM adjuvant containing Quillaja saponaria saponins.Materials and methods. The Matrix-V ISCOM adjuvant was produced using Vector’s proprietary technology, which involves cross-flow filtration through Sartorius VivaFlow cassettes. To determine the saponin and residual detergent concentrations in Matrix-V, the authors conducted high-performance liquid chromatography. Having produced the recombinant SARS-CoV-2 RBD antigen, the authors used electron microscopy to analyse the ultrastructure of the ISCOM–antigen complex. In the study of the ISCOM–antigen complex, 25 female Balb/c mice (5 groups) and 15 male and female outbred guinea pigs (3 groups) received two intramuscular injections with a 14-day interval. Serum tests relied on virus neutralisation (VN) and enzyme-linked immunosorbent assay (ELISA) methods and used antigens of 8 SARS-CoV-2 variants (State Collection of Viruses and Rickettsia, Vector). The authors used Statistica 10 to analyse the results.Results. Two injections of the SARS-CoV-2 RBD antigen (mice: 7 μg, guinea pigs: 1 μg) alone did not induce statistically significant virus-neutralising antibody responses, as shown by the VN results. Two injections of the SARS-CoV-2 RBD antigen (mice: 7 μg, guinea pigs: 1 μg) adjuvanted with Matrix-V (25 μg) resulted in geometric mean antibody titres of 1:83–1:178 (mice) and 1:174–1:587 (guinea pigs) in the VN tests with the Wuhan variant. One injection of the antigen (1 μg or 7 μg) with Matrix-V (25 μg) induced antibodies only in individual cases, as demonstrated by the VN and/or ELISA results. The most intensive immune response was observed in ELISA tests with the Delta variant after two injections of the Ecto-S-Wuhan (1 μg) and Matrix-V (25 μg) complex. Immune responses did not differ between the group that received two injections of the Ecto-S-Wuhan antigen (1 μg) without the ISCOM adjuvant and the negative control group (titres below 1:100; p=0.95). Two injections of the SARS-CoV-2 RBD antigen (7 μg) without the ISCOM adjuvant induced antibodies in mice (titres between 1:248 and 1:1477).Conclusions. Two intramuscular injections of the complex containing the recombinant SARS-CoV-2 RBD antigen and the Matrix-V ISCOM adjuvant induce virus-neutralising antibodies. The approach proposed by the authors has the potential for use in the development of immunobiological medicinal products to prevent and treat a wide range of infectious diseases.

Comparative Analysis of UV and UV-LED Light Traps for Pest Control: A Cost-Efficiency Perspective

This research, conducted between October 2019 and March 2020 at the Biotechnology Centre, Jawaharlal Nehru KrishiVishwaVidyalaya, Jabalpur, Madhya Pradesh, aimed to compare the effectiveness and cost-efficiency of UV and UV-LED light traps in attracting and capturing insect pests. The study utilized UV 15-watt (model SMV-4) and UV LED 7-watt solar trap (model Rakshak) to assess their respective performances in capturing Gryllusbimaculatus, unidentified Lepidoptera moths, Helicoverpaarmigera, Gryllotalpaorientalis, Plusiaorichalcea, Agrotisipsilon, and Creatonotosgangis. The results revealed that the UV 15-watt trap demonstrated superior efficacy in capturing Gryllusbimaculatus and unidentified Lepidoptera moths, while no significant difference was observed between UV 15W and 7W UV LED traps.Furthermore, the UV 15-watt trap outperformed the UV LED 7-watt in capturing Helicoverpaarmigera, Gryllotalpaorientalis, Plusiaorichalcea, Agrotisipsilon, and Creatonotosgangis, with significant differences noted in the catches. Despite these variations, considering the total wattage consumption, the UV 7-watt solar trap emerged as a more cost-effective alternative, showcasing economic advantages over the UV 15-watt electric-powered counterpart. Conclusively, the solar-powered UV 7-watt light trap proves to be a promising substitute for the UV 15-watt model in pest control applications. However, it is crucial to acknowledge the higher initial cost associated with the solar-powered light trap, highlighting a potential trade-off between cost and efficiency in light trap selection for pest management.

Molecular Detectıon of Canine Leishmaniasis in Northern Anatolia, Turkiye.

Canin leishmaniasis (CanL), mostly caused by Leishmania infantum, is one of the most important vector-borne diseases in dogs in the Mediterranean region. In this study, we aimed to determine the disease profile in this region by firstly making microscopic and then molecular analyzes in the samples taken from the dogs. Overall, 112 whole blood samples taken from dogs for clinical applications by a veterinarian in Cankırı between December 2021 and November 2022 were used. After blood collection, both thin and thick drop blood smear preparations were prepared and evaluated for Giemsa staining. L. infantum was investigated by Real time-PCR (RT-PCR) method from all blood samples. Sequence analysis and phylogenetic tree study were performed on positive samples. Both microscopic and RT-PCR analyzes were performed. In both studies, 3 of the 112 samples were positive. Because of the sequence analysis, they were L. infantum. Sequence analysis was performed from the samples found 3 positive. The phylogenetic tree was drawn by making NCBI (National Center for Biotechnology) data entries of the positive samples (Accession numbers: OQ184728, OQ184729, OQ184730). Dogs are important, as they are reservoir of this disease. In this study, 3 (2.7%) positive Leishmaniasis was detected in dogs in Cankırı. Ultimately, this should prompt discussion about new strategies going forward to combat infection caused by Leishmania.

Сравнительная оценка токсичности и вирулицидных свойств дезинфектантов, применяемых при работе с вирусом SARS-CoV-2 в изолированных вирусологических лабораториях

The choice of disinfectants and their combinations for the final disinfection of the premises of «infectious» areas of laboratories of biosafety level 3–4, in which work is carried out with viruses with epidemic potential, is important for reducing the toxic effect of these disinfectants on personnel while maintaining 100?% disinfection efficiency. Purpose of the study: to study the toxicity of disinfectants of different groups for cell culture, to select a toxicity neutralizer, to study the virucidal activity of these disinfectants and their combinations against the SARS-CoV-2 virus. Materials and methods: experimental studies were carried out using the reference strain of coronavirus SARS-CoV-2, namely the hCoV-19/Australia/VIC01/2020 (AUS) strain (Wuhan variant), obtained from the State Collection of Causative Agents of Viral Infectious Diseases and Rickettsial Diseases, functioning on the basis of the Federal Budgetary Institution State Scientific Center for Virology and Biotechnology «Vector» of Rospotrebnadzor. The residual infectious activity of the virus was determined by titrating samples in cell culture. To conduct the research, we used a Vero E6 cell culture obtained from the Collection of Cell Cultures of the Federal Budgetary Institution of Scientific Research Center for Virology and Biochemistry «Vector» of Rospotrebnadzor in the form of a 2-day monolayer with a confluency of 95–100?%. Results. As a result of the studies, experimental data were obtained on the dynamics of the virucidal activity of different groups of disinfectants and their combinations against the SARS-CoV-2 virus. The degree of toxicity of disinfectants for Vero E6 cell culture is shown. Optimal combinations of disinfectants and a 40?% aqueous solution of formaldehyde (hereinafter referred to as formalin) have been selected, increasing its disinfection efficiency at an initial concentration halved, which can be used for final disinfection treatment of the premises of isolated virology laboratories conducting work with the SARS-CoV- coronavirus. Conclusion. The results obtained indicate that when using selected combinations of disinfectants to carry out final disinfection with formaldehyde vapors formed during the evaporation of formaldehyde as a result of its heating, a twofold decrease in the initial concentration of formalin is possible, leading to a decrease in its toxic effect on personnel, while maintaining 100?% efficiency disinfection. Keywords: disinfection, disinfectants, formaldehyde, SARS-CoV-2 coronavirus, disinfection efficiency, cytotoxicity.

Formation of the All-Russian Education System in the field of healthy food of the population

The establishment of the educational cluster “Healthy Eating” in the Russian Federation is based on the Federal State Budgetary Institution’s “Federal Research Center for Nutrition and Biotechnology” and intended to save the health of children and adults, as it serves as a model for the children’s and adults’ population. The development of different programs for both specialists and the population has been done through various means. It is suggested to concentrate attention on two main areas of activity: educational and outreach. Learning activity in the form of a cluster is shown to create the conditions for the development of an all-Russian system of education in the field of healthy nutrition for the Russian Federation’s population.
 Aim. Development and implementation of a system of federal and regional educational programs for specialists and various groups of the population of the Russian Federation.
 The population of the Russian Federation has an insufficient level of knowledge about healthy nutrition, so about 50% have practically no idea about its rules, 60% of the population do not follow the diet. In connection with the urgency of the problem, an educational cluster was formed on the basis of the Federal Research Center for Nutrition and Biotechnology. The resources and qualifications of the cluster members, which combined the scientific potential of the Federal Research Center for Nutrition and Biotechnology and the educational opportunities of specialized departments of the country’s leading universities, make it possible to effectively solve the problems that the education system of specialists and the population faces in the field of healthy nutrition. Additional education programs have been developed – advanced training courses, differentiated programs are being created for specialists: nutritionists, doctors, faculty of medical universities, teachers and trainers of sports schools, fitness trainers, food industry workers, graduate students and residents, as well as educational and educational programs for various population groups.
 Taking into account the high efficiency of outreach work, as well as its relatively low cost, in addition to the educational block, an information and educational block has been allocated separately, including with the involvement of public opinion leaders and the media.
 Thus, the establishment of a systematic approach to the education of specialists and the public in the field of healthy nutrition, through management, scientific, methodological, and educational support within the educational cluster, leads to the creation of an all-Russian system of education in healthy nutrition for the Russian population.
 Conclusion. Successful implementation of federal and regional programs of the educational cluster “Healthy Nutrition” is a necessary condition for improving the quality of life and health of the population.

Evaluation for Parental Polymorphism and Identification of Microsatellites Linked to Drought Tolerance in Rice (Oryza sativa L.)

Aims: To identify the ready-to-use polymorphic microsatellite markers associated with drought tolerance for marker-assisted backcross breeding through a polymorphism survey between Jyothi and Chuvannamodan rice varieties.
 Place and Duration of Study: Centre for plant biotechnology and molecular biology, Thrissur, Kerala, India, during January to May 2023.
 Methodology: The genomic DNA of Jyothi and Chuvannamodan was isolated by following CTAB (Cetyltrimethyl Ammonium Bromide) method with modification. Isolated DNA from both varieties was subjected to PCR amplification using 208 Simple Sequence Repeats (SSRs) primer pairs distributed in 12 chromosomes. The amplified PCR products were electrophoresed in 3% agarose gel and separated fragments were visualized and documented in the Gel Documentation System. Different allele sizes produced by the same SSRs primers between two varieties are identified as polymorphic markers. Polymorphism per-cent was calculated, and frequency distribution and chromosome distribution of polymorphic markers were analyzed.
 Results: A total of 208 SSR primers were surveyed for the parental polymorphism. Out of which, 85 SSR primers exhibited clear polymorphism between Jyothi and Chuvannamodan and the remaining 123 were monomorphic primers. The amplicon size ranged from 83bp (RM430) to 495bp (RM18919) among the different primers. The survey revealed maximum parental polymorphism on chromosome 4 (69.23%), followed by chromosome 5 (64.28%), and minimum polymorphism on chromosome 8 (21.73%). The average per cent of polymorphism between the parents was 40.86%. Among the 85 polymorphic markers, 66 had dinucleotide repeats, 17 had trinucleotide repeats, and 1 had tetranucleotide repeats.
 Markers with dinucleotide repeats showed higher level of polymorphism. A group of 28 polymorphic markers were identified to be linked with traits including root-related traits, grain yield, leaf rolling and leaf drying under drought conditions.
 Conclusion: The polymorphic markers identified in the present study form the basis for tagging drought-tolerant QTLs/genes, fine mapping of those genes, and subsequently in marker-assisted breeding programs. The polymorphic markers linked with the QTLs for grain yield under drought can be pyramided in Jyothi through marker-assisted backcross breeding.

First person – Kolaparamba V. Navyasree

ABSTRACT First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping researchers promote themselves alongside their papers. Kolaparamba V. Navyasree is first author on ‘ Cholesterol regulates insulin-induced mTORC1 signaling’, published in JCS. Kolaparamba is a PhD student in the lab of Perunthottathu K. Umasankar at Rajiv Gandhi Centre for Biotechnology (RGCB), Trivandrum, Kerala, India, where she explores the intricate mechanisms of cell signaling pathways to unravel their role in cellular function and potential diseases.

The improvement of the in vitro plant regeneration in barley with the epigenetic modifier of histone acetylation, trichostatin A

Genotype-limited plant regeneration is one of the main obstacles to the broader use of genetic transformation in barley breeding. Thus, developing new approaches that might improve responses of in vitro recalcitrant genotypes remains at the center of barley biotechnology. Here, we analyzed different barley genotypes, including “Golden Promise,” a genotype commonly used in the genetic transformation, and four malting barley cultivars of poor regenerative potential. The expression of hormone-related transcription factor (TF) genes with documented roles in plant regeneration was analyzed in genotypes with various plant-regenerating capacities. The results indicated differential expression of auxin-related TF genes between the barley genotypes in both the explants and the derived cultures. In support of the role of auxin in barley regeneration, distinct differences in the accumulation of free and oxidized auxin were observed in explants and explant-derived callus cultures of barley genotypes. Following the assumption that modifying gene expression might improve plant regeneration in barley, we treated the barley explants with trichostatin A (TSA), which affects histone acetylation. The effects of TSA were genotype-dependent as TSA treatment improved plant regeneration in two barley cultivars. TSA-induced changes in plant regeneration were associated with the increased expression of auxin biosynthesis-involved TFs. The study demonstrated that explant treatment with chromatin modifiers such as TSA might provide a new and effective epigenetic approach to improving plant regeneration in recalcitrant barley genotypes.

Candida albicans and Antifungal Peptides.

Candida albicans, a ubiquitous opportunistic fungal pathogen, plays a pivotal role in human health and disease. As a commensal organism, it normally resides harmlessly within the human microbiota. However, under certain conditions, C.albicans can transition into a pathogenic state, leading to various infections collectively known as candidiasis. With the increasing prevalence of immunocompromised individuals and the widespread use of invasive medical procedures, candidiasis has become a significant public health concern. The emergence of drug-resistant strains further complicates treatment options, highlighting the urgent need for alternative therapeutic strategies. Antifungal peptides (AFPs) have gained considerable attention as potential candidates for combating Candida spp. infections. These naturally occurring peptides possess broad-spectrum antimicrobial activity, including specific efficacy against C.albicans. AFPs exhibit several advantageous properties, such as rapid killing kinetics, low propensity for resistance development, and diverse mechanisms of action, making them promising alternatives to conventional antifungal agents. In recent years, extensive research has focused on discovering and developing novel AFPs with improved efficacy and selectivity against Candida species. Advances in biotechnology and synthetic peptide design have enabled the modification and optimization of natural peptides, enhancing their stability, bioavailability, and therapeutic potential. Nevertheless, several challenges must be addressed before AFPs can be widely implemented in clinical practice. These include optimizing peptide stability, enhancing delivery methods, overcoming potential toxicity concerns, and conducting comprehensive preclinical and clinical studies. This commentary presents a short overview of candidemia and AFP; articles and reviews published in the last 10years were searched on The National Library of Medicine (National Center for Biotechnology Information-NIH-PubMed). The terms used were C.albicans infections, antimicrobial peptides, antifungal peptides, antifungal peptides mechanisms of action, candidemia treatments and guidelines, synthetic peptides and their challenges, and antimicrobial peptides in clinical trials as the main ones. Older publications were cited if they brought some relevant concept or helped to bring a perspective into our narrative. Articles older than 20years and those that appeared in PubMed but did not match our goal to bring updated information about using antifungal peptides as an alternative to C.albicans infections were not considered.

PSIV-30 Exploring the Prebiotic Activities of Proanthocyanidins on a Platform Using the Three-Dimensionally (3D)-Cultured Organoids

Abstract Using antibiotics, the antimicrobial substance active against bacteria and promoting growth efficiency, in feedstuffs and food supply has been attracting great attention due to its side effects and contribution to antibiotic resistance. Therefore, exploring talternatives to antibiotics is of great significance and urgent need for human health and animal industries with a sustainable high efficiency. The objective of this study is to evaluate the biological and medicinal activities of the monomers of proanthocyanidins (PAs), such as epicatechin, epigallocatechin gallate, and flavanols such as quercetin, isoquercetrin, and rutin generated from engineered plants. The evaluations were performed in intestinal organoids isolated from ileum of neonatal piglets. The organoids after expended in vitro were incubated with or without the candidate compounds for 24 hours, and then treated with or without polyinosinic-polycytidylic acid [Poly (I:C), 10 µg/mL] for another 24 hours, a synthetic analog of double-stranded RNA (can induce a molecular pattern associated with viral infections). Cell proliferation, barrier function, toll-like receptor pathway and apoptosis as well as organic cation transporters were assessed by measuring the abundance of the corresponding genes. Concentration gradient (0 to 200 µM) measurements showed that isoquercetrin and epigallocatechin gallate might stimulate the intestinal cell proliferation, promote the uptake of L-carnitine, and increase barrier function and mucin secretion (P < 0.05). Epigallocatechin gallate and epicatechin might affect the inflammation responses via modifying expression of Interleukins but not induce apoptosis. Interactions between the examined these compounds and Poly (I: C) were not observed (P > 0.05), but the influence on the challenged pattern were tested for some of the measured genes (P < 0.05). Based on the data collected from this in vitro study, we conclude that epigallocatechin gallate is a potential PA monomer with all prebiotic characteristics and its applicable values in feedstuff and food supply should be studied in vivo studies in domestic (food) animals with and without antigen challenges. Supported by NC Biotechnology Center project (1107)2022-3082 and the North Carolina Agricultural Research Hatch Projects 02780.

Diagnosis of two local mushroom species (Pleurotus spp.) and their production management

The present study confirmed the identity of two Iraqi mushrooms by using the internal transcribed spacer (ITS) sequence; the genomic DNA of two isolates was amplified using ITS1 and ITS4 primers, and the data analyzed through Basic Local Alignment Tool( BLAST) search was achieved using the National Center for Biotechnology Information (NCBI) database. The result shows that the nucleotide sequence of two mushrooms blasted against the sequence from the Gene Bank database that Pleurotus ostreatus and Pleurotus eryngii matched 99% and 100%, respectively. The second stage of the current study was the comparison of different agro-waste, including wheat straw(WS), corncob(CC) and sawdust(SD) supplemented with 25% wheat bran(W) and 2% calcium carbonate (CaCo3) on mycelium growth, yield and biological efficiency (BE) of local oyster mushroom which comparison with Chinese strain pleurotus ostreatus121 and pleurotus eryngii 080. The highest growth average of local P. ostretus on the SD substrate was 1.50 cm, and P.eryngii was recorded at 1.18 cm. The substrate CC was the most suitable for the yield of mushroom local P. ostreatus and P. ostreatus121(485.40 and 418.50 g/bag, respectively), while local P. eryngii recorded 470.40 in the same substrate. It was also found that the WS substrate recorded the highest BE for each local isolate and Chinese strain. Keywords: Iraqi strains, Pleurotus spp., agricultural wastes, wild agricultural mushrooms

FACTORS THAT INFLUENCE THE PREGNANCY RATE IN BIOTECHNOLOGIES APPLIED ON RURAL PROPERTIES IN THE WEST GOIANO LÁCTEO APL

The present study sought to investigate the effect of the degree of the corpus luteum, how the inovulation was performed (left or right) and season of the year related to forage availability, on the pregnancy rate of crossbred heifers submitted to TETF (embryo transfer in fixed time). This is a retrospective study that analyzed data recorded by BIOTEC (Center for Biotechnology in Animal Reproduction) belonging to the State University of Goiás, West Campus, located in São Luís de Montes Belos. The records had records of the diagnosis of pregnancy, inovulated horn and degree of corpus luteum of 206 females who received cryopreserved embryos from IVF. The transfers were carried out between February and August 2021. The data were represented in tables and later analyzed and discussed, in order to assess the effect of the aforementioned factors on the pregnancy rate of recipients in this analyzed group. Testing the hypotheses of several authors, it was verified: The degree of CL did not influence the pregnancy rate at 30 days of the recipients; The pregnancy rate at 30 days in recipients was higher in inovulations performed in the left horn, adjacent to the left ovary with the presence of CL; The pregnancy rate was higher in autumn compared to winter and summer.

Screening of antiviral activity of samples from chaga Inonotus obliquus and humic acid from brown coal on Vero cell culture against ectromelia virus (Poxviridae: Orthopoxvirus; ECTV)

The mouse-specific orthopoxvirus, ectromelia virus, is one of the best models that can be used to study key issues of pathogenesis, prevention, and treatment of smallpox, and to develop measures to increase virulence, transmissibility, or the ability to overcome vaccine immunity. The aim of the work is to screen the antiviral activity of samples from Inonotus obliquus chaga and humic acid from brown coal in vitro against ectromelia virus. We used ectromelia virus, strain K-1 (reg. No V-142), obtained from the State Collection of Pathogens of Viral Infections and Rickettsioses of the State Scientific Center of Virology and Biotechnology "Vector"; Vero Е6 cell culture (No 70) from the Collection of cell cultures of the State Scientific Center of Virology and Biotechnology "Vector". Nine samples from chaga I. obliquus and humic acid from brown coal were used to evaluate the changes in the infectivity of the ectromelia virus on cell culture using 2 schemes of application of drugs and virus (preventive and therapeutic schemes), and to assess their cytotoxicity and antiviral activity. 50% cytotoxic concentration, 50% virus-inhibiting concentrations and selectivity index were determined for all samples. The studied samples were shown to be non-toxic to the monolayer of Vero cell culture in a dilution of 300 and more micrograms/ml, while demonstrated high antiviral activity against strain K-1 of ectromelia virus in two application schemes - preventive and curative. All samples tested for ectromelia virus in vitro can be considered promising for further development of drugs against diseases caused by orthopoxviruses.