Biotechnologia最新文献

Background: Conotoxins are small peptides known for their potent and selective activity on ion channels, offering potential applications in both medicine and cosmetology. This study aimed to design and validate recombinant conotoxin TIIIA and its mutant TIIIAlaMut, assess their biological activity on the voltage-gated Na⁺ (Nav) channel Nav1.4, and evaluate the antiwrinkle efficacy of a topical cream containing the recombinant peptide in a group of volunteers.

Materials and methods: Fusion genes encoding TRX::TIIIA and TRX::TIIIAlaMut were cloned into the pDM vector and expressed in Escherichia coli S4B cells. The proteins were purified using Ni-NTA chromatography, cleaved with CNBr under optimized acidic conditions, and analyzed. Biological activity was assessed using two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing the human Nav1.4 channel. Additionally, a conotoxin-containing cream was applied to 55 human volunteers in an application study assessing its antiaging effects.

Results: Both recombinant genes were successfully expressed, purified, and activated. Electrophysiological measurements demonstrated their ability to inhibit Nav1.4 channel activity, including the version extracted directly from the cream. In the human study, 47% of participants reported a visible reduction in wrinkles. Additional benefits included evening of skin tone, reduced erythema, and balanced sebum production in oily skin types.

Conclusion: This study describes the design, bacterial expression, and functional analysis of recombinant conotoxins TIIIA and TIIIAlaMut. Their bioactivity was confirmed on human Nav1.4 channels. The recombinant toxins, including the form extracted from the cream, showed effects comparable to a synthetic standard. Application tests demonstrated the conotoxin's potential in cosmeceuticals, particularly in reducing periocular wrinkles and improving skin texture and tone.

Background: As society rapidly ages, the escalating global demand for natural, high-value antioxidants - particularly ketocarotenoids such as canthaxanthin - is driving intensive research into their sustainable bioproduction. This study investigates the potential of the green microalga Chlorosarcinopsis PY02 as a novel microbial cell factory for enhanced ketocarotenoid production under abiotic stress conditions.

Materials and methods: We optimized bioprocess parameters using a simple, spot-test-based high-throughput screening technique, evaluating algal growth and pigment accumulation on tris acetate phosphate agar supplemented with various sodium chloride concentrations (0-15 g/l).

Results: Peak canthaxanthin content (294.55 µg/g dry weight) was observed at 10 g/l NaCl, while biomass yield was highest at 12 g/l. Combining salt stress with a 50% nitrogen reduction increased total carotenoid productivity (highest with 10 g/l NaCl: 3.10 mg/l) but did not enhance canthaxanthin levels; the salt-only treatment produced the highest canthaxanthin yield (0.80 mg/l). Pigment identification and quantitative profiling were performed using thin-layer chromatography and spectrophotometry, confirming the efficiency of the production process.

Conclusions: These findings highlight Chlorosarcinopsis PY02 as a promising candidate for sustainable, large-scale production of ketocarotenoids. The study also demonstrates a cost-effective and scalable approach for inducing carotenoid biosynthesis in Chlorosarcinopsis PY02, with strong relevance for sustainable pigment production. The simple visual screening method provides a practical tool for preliminary strain and condition optimization in microalgal bioprocess development.

Background: Polycystic ovary syndrome (PCOS) affects millions of women worldwide and is primarily known for its reproductive and hormonal symptoms. However, growing evidence suggests a strong link between PCOS and inflammation. Rheumatoid arthritis (RA) and osteoarthritis (OA) similarly involve systemic inflammation and immune dysregulation. Despite their distinct clinical manifestations, these disorders may share overlapping biological pathways. This study aimed to identify shared transcriptomic signatures between PCOS and autoimmune joint diseases such as RA and OA.

Materials and methods: RNA sequencing datasets were downloaded from the publicly available Gene Expression Omnibus (GEO) database. After processing and quality filtering, a total of 73 samples from the GSE277906 and GSE89408 datasets were selected. DEG analysis was conducted using the DE-Seq2 package in RStudio by adjusting for a significant p-value < 0.1 and |log₂ fold change| > 0.5. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to determine functional enrichment of genes and common pathways associated with the diseases.

Results: A total of 10,492 and 9,892 DEGs were identified in PCOS vs. RA and PCOS vs. OA, respectively. Key genes dysregulated among the diseases included TOMM34, DHCR24, CMAS, RBP1, and HSD3B2, and the enrichment analysis revealed overlapping pathways involving immune regulation, mitochondrial dysfunction, oxidative stress, and proteasome activity. Notably, 201 GO pathways were shared by PCOS and OA, 123 by RA and OA, and 267 by PCOS and RA. All three conditions shared a set of 57 GO pathways, including mitophagy and ER stress.

Conclusion: The identified common pathways signify the overlap between PCOS, RA, and OA. These findings support the hypothesis of systemic immunometabolic involvement in PCOS.

Molecular profiling has become a cornerstone of cancer diagnosis and treatment, with BRAF alterations serving as significant markers across various tumor types. The gene encodes a serine/threonine kinase involved in the MAPK/ERK signaling pathway, which regulates cell proliferation and survival. Mutations in BRAF, notably the V600 codon substitutions, are among the most common genetic drivers in melanoma and other cancers, including thyroid, colorectal, and non-small cell lung cancer. BRAF mutations are categorized into three functional classes (class I-III), each with distinct activation mechanisms and therapeutic implications. Current targeted therapies - primarily BRAF and MEK inhibitors, including the first FDA-approved anti-BRAF tumor-agnostic therapy - are most effective in cancers harboring the class I V600E mutation. However, the emergence of resistance to BRAF inhibitors has driven the development of next-generation inhibitors and combination treatments. Furthermore, innovative immunotherapy-based treatments have demonstrated synergistic potential in specific BRAF-mutated malignancies. Accurate molecular diagnostics are crucial in cancer treatment; therefore, numerous molecular diagnostic methods are employed, including next-generation sequencing (NGS), quantitative PCR, droplet digital PCR, Sanger sequencing, and fluorescence in situ hybridization (FISH). NGS, particularly comprehensive genomic profiling, provides the broadest and most detailed genetic data, although simpler laboratory techniques remain popular due to their accessibility and straightforward protocols. Further research into resistance mechanisms and combination therapies, as well as the integration of circulating tumor DNA (ctDNA) in diagnostics, is needed to fully realize the potential of personalized treatment in BRAF-driven tumors.

Background: Soil contamination by oil products is a significant problem that affects the environment, agriculture, economy, and human health, and requires effective solutions. The study aimed to develop effective methods of bioremediation of oil-contaminated soils using microbial preparation D (a mixture of Rhodococcus sp. and Gordonia sp. - a consortium of autochthonous hydrocarbon-degrading micro-organisms), a rhamnolipid biocomplex (RBC), the oxidant calcium peroxide (CaO₂), and plant remediants.

Materials and methods: Bioremediation processes were carried out on oil-contaminated clay soil (initial contamination - 9.5%) over 1.5 years. First, the soil was treated with microbial preparation D and CaO₂. After 14 days, field peas or sorghum were sown, with seeds treated using an RBC solution. Hydrogen peroxide content and lipid peroxidation index in plants, as well as soil dehydrogenase activity, were determined by spectrophotometry. Additionally, soil phytotoxicity was assessed using test plants, and the residual content of oil products was quantified.

Results: The best effect was achieved with the combined use of microbial preparation D, RBC, and CaO₂: the degree of oil contamination in the soil decreased to 1.3%; with microbial preparation D, plants, and RBC, contamination decreased to 1.4-1.6% (compared to the initial 9.5%). The maximum value of dehydrogenase activity was recorded when sorghum, microbial preparation D, and RBC were applied, 2.7 times higher than in the control. After bioremediation, the phytotoxicity of oil-contaminated soils (in test plants) decreased on average by 3.7 times compared to the control.

Conclusion: The effectiveness of the integrated use of hydrocarbon-degrading microorganisms, field peas, sorghum, RBC, and CaO₂ in bioremediation of oil-contaminated soils was established.

Nanotechnology has emerged as a promising field with the potential to revolutionize several industries, including the food industry. It offers innovative solutions to critical challenges in food, such as safety, nutrition, waste reduction, and sustainability. This study examines the possibilities offered by nanotechnology in the food sector, with a focus on risk assessment, safety evaluation, and regulatory approaches. While nanotechnology in food applications presents many advantages, it also raises concerns about potential health risks. Due to their distinct characteristics, nanoparticles may interact with living organisms in unpredictable ways, creating challenges for risk assessment and management. This review also explores the possible hazards of using nanomaterials in the food system, highlighting the need for comprehensive toxicity studies and effective regulatory frameworks. Addressing these issues requires a multidisciplinary approach involving collaboration among scientists, regulators, policy-makers, and stakeholders to balance the benefits and risks of nanotechnology in the food system. As the food sector seeks novel approaches to meet rising global demand, it is crucial to thoroughly assess both the advantages and risks of nanotechnology to ensure its responsible and sustainable application while protecting human health and the environment.

Background: Changes in biomolecules under the influence of chemical and physical factors on cells, tissues, and whole organisms are investigated.

Materials and methods: In vitro-incubated bovine embryonic cells were inoculated with low titers (high dilutions of viral suspensions) of vaccine avipoxviral strains. Mouse embryonic fibroblasts were co-cultivated with mouse malignant myeloma cells (P3-X63-Ag8) transfected by recombinant DNA plasmid or preincubated in culture fluid from prior incubation of the same cells. Sub-populations of virus-inoculated, co-cultivated, and preincubated cell cultures were frozen in the presence of the cryoprotectant dimethylsulfoxide (DMSO), subsequently thawed, and re-incubated. Newly formed cell monolayers were inoculated with extracellular and intracellular forms of each viral strain, both before and after exposure to DMSO and drastic temperature changes. Extracellular forms were derived from the cultural fluids of inoculated cell cultures, while intracellular forms were obtained from suspensions of mechanically scraped virus-inoculated cells.

Results: Exchange of nucleotide (DNA and/or RNA) fragments between cellular and viral genomes, as well as between genomes of separate cells, was suggested. These changes were explained by activated fusion induced by the organic detergent (DMSO) combined with drastic temperature changes. Such processes could provide vectors for gene-engineering manipulations and the development of molecular (DNA-based, RNA-based, and/or protein-based) antiviral and antimalignant vaccines. Production of immune molecules by nonimmune cell types under appropriate conditions, such as the presence of immunomodulators, was also proposed.

Conclusions: The results suggest the possibility of nucleotide (DNA and/or RNA) fragment exchange between separate cells, as well as between cells and virions. Nonimmune cells demonstrated the capacity to produce immune molecules under appropriate conditions.

Background: Synthetic colors are widely used in the food and cosmetic industries to make products more appealing to consumers. However, the health hazards associated with synthetic colors have prompted their replacement with natural colors. Portulaca grandiflora is a promising candidate for natural color extraction, as it is rich in betalains. This study presents a reliable, reproducible three-step regeneration protocol for this plant and analyzes its betalain content.

Materials and methods: In vitro shoot cultures were established on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. Anatomical studies were conducted to determine the stages of shoot primordium development. Betalains were extracted from in vivo plants using 60% methanol and subjected to spectrophotometric analysis. The effect of sodium ascorbate on betalain stability was also evaluated.

Results: Juvenile leaf explants regenerated shoots on MS medium supplemented with 10 μM 6-benzyladenine and 5 μM indole-3-acetic acid. Shoots were multiplied with 20 μM BA (6.25 ± 0.85 shoots/explant), and elongation was achieved with 5 μM gibberellic acid (GA₃) (8.2 ± 0.37 shoots/explant). Shoot primordia developed from well-organized meristemoid cells. The betalain content in the stem was 26.66 ± 0.19 mg/100 g, but this pigment degraded within 24 h (42.19% degradation). The addition of 50 mM sodium ascorbate prevented betalain degradation, even after 24 h.

Conclusion: This study reports a regeneration protocol from juvenile leaf explants and demonstrates that betalain stability in the stem can be maintained with 50 mM sodium ascorbate.

Background: Helicobacter pylori infects approximately half of the global population, leading to gastric and duodenal ulcers. Despite the availability of antibiotics, challenges such as patient reluctance, high treatment costs, and antibiotic resistance limit their effectiveness, making vaccination a promising alternative. This study used immunoinformatics to identify candidate epitopes for a multiepitope vaccine construct against H. pylori.

Materials and methods: The protein variability server was utilized for conservation analysis. The epitopes were screened for antigenicity, allergenicity, toxicity, cross-reactivity, and population coverage. Selected epitopes were docked with their corresponding human leukocyte antigen (HLA) alleles, and thermodynamic quantities were determined. Five virulence factors - HopZ, SabA, HP-NAP, OipA, and urease - were selected for their critical roles in bacterial adhesion, immune modulation, and stress survival.

Results: Conservation analysis revealed a highly conserved protein sequence (Shannon index ≤ 0.1). The predicted epitopes had an IC₅₀ value of ≤ 500 nM, indicating strong binding to the corresponding HLAs, with an estimated population coverage of more than 90% in the Southeast Asian region. The predicted epitopes were identified as probable nonallergens, nontoxic, and noncross-reactive (E value >1.0). Molecular docking analysis showed that the candidate epitopes could bind strongly and spontaneously with their corresponding HLA proteins, as evidenced by low negative Gibbs free energy (ΔG) values and dissociation constants (K _D < 100 nM).

Conclusion: The epitopes predicted from the five virulence factors present promising candidates for future H. pylori vaccine design. Further in vitro and in vivo experiments are recommended to validate these preliminary findings.

In this review, I describe recent findings on the molecular architecture and genomic characterization of giant viruses that infect microbial eukaryotes (protists) across diverse ecosystems and ecological niches. Giant viruses are distinguished by their large and complex genomes, which encode a wide range of functions, including protein translation, carbohydrate and lipid metabolism, nitrogen cycling, light assimilation, and key metabolic pathways such as glycolysis and the tricarboxylic acid cycle. Additionally, these genomes feature unique genes, often acquired through horizontal gene transfer, that are not found in other viruses and contribute to the viruses' ability to manipulate host metabolism and evade host defenses. A core set of genes conserved across different families of giant viruses is highlighted, serving as essential components for key life-cycle processes and providing valuable phylogenetic markers. The review also discusses the role of ORFans and virophages in contributing to the genetic diversity and evolutionary adaptation of these viruses. These findings are crucial for understanding the diversity, evolutionary mechanisms, and complex virus-host interactions of giant viruses, as well as for developing more advanced classification systems. Furthermore, the potential biotechnological applications of unique viral genes and pathways are explored, underscoring the importance of ongoing research in this field.