Biotechnology and applied biochemistry最新文献

Thiourea structures, known for their wide-ranging bioactivity, have significant potential in diabetes management. In this study, it was aimed to examine the antioxidant capacities of fluorophenyl thiourea derivative compounds and their inhibition studies on α-amylase and α-glycosidase enzyme activity. Antioxidant capacity was determined using Fe³⁺-Fe⁺², FRAP, and Cu²⁺-Cu⁺ reducing analyses, DPPH· and ABTS·⁺ scavenging experiments. It was observed that fluorophenyl thiourea derivative compounds exhibited quite high antioxidant activity compared to standard antioxidants such as BHA, BHT, trolox, α-tocopherol, and ascorbic acid. Additionally, this study investigated the inhibitory effects of the analysis molecules on α-glycosidase and α-amylase, which are enzymes associated with diabetes. Among these derivative molecules, 4-fluorophenyl showed the highest inhibition on α-amylase (IC₅₀: 53.307 nM) and α-glycosidase (IC₅₀: 24.928 nM). These results highlight the potential of thiourea derivatives in enzyme inhibition and antioxidant therapy, making them promising candidates for diabetes management.

Small molecule targeted inhibitor therapies often have several drawbacks, including limited oral bioavailability, quick metabolism, toxic effects that limit dosage, and poor water solubility. This study aims to develop a nanodrug self-delivery system that does not require a carrier by utilizing the self-assembly of camptothecin (CPT) and dihydroartemisinin (DHA). CPT/DHA nanoparticles (NPs) with varying diameters can be synthesized without requiring further carrier materials or chemical modifications by changing the CPT-to-DHA ratio (10:1, 5:1, 2:1, 1:1). Even more crucially, CPT/DHA NPs generate an AIE impact when they self-assemble. CPT/DHA NPs are used for cell tracking and bioimaging fluorescent probes. We chose CPT/DHA NPs (2:1) with a size of approximately 140 nm for the anticancer examinations. The A549 cells were used to assess the cytotoxicity, morphological changes by biochemical staining methods and apoptosis by flow cytometric techniques of CPT/DHA NPs. Finally, in vitro anticancer research proved that CPT/DHA NPs are biocompatible and have strong synergistic anticancer properties.

Magnetic nickel oxide multi-walled carbon nanotubes (MWCNT-NiO) were employed in the immobilization of xylanase from Thermomyces lanuginosus, after modification with (3-glycidoxypropyl)trimethoxysilane or 3-aminopropyltriethoxysilane (APTES). The APTES-derivatized MWCNT-NiO particles were activated with glutaraldehyde to immobilize T. lanuginosus xylanase via covalent attachment. The (3-glycidoxypropyl)trimethoxysilane-derivatized MWCNT-NiO particles were directly used for the covalent immobilization of T. lanuginosus xylanase, or the formed epoxy groups were converted to aldehyde groups. The free xylanase had maximum activity at pH 7.5, whereas the immobilized samples showed an optimum pH of 7.0. The optimum temperature was 60°C for the xylanase samples. The thermal stability of xylanase increased at 7 and/or 12 folds after immobilization. The results of xylooligosaccharide synthesis showed that the main formed xylooligosaccharides were xylobiose, xylotriose, and xylotetraose for the immobilized xylanase samples. Furthermore, an effect of the enzyme loading could be found, an increase in this parameter promoted that xylobiose and xylotriose amounts slightly increased, whereas xylotetraose amount slightly decreased. The immobilized xylanase samples retained at least 80% of their initial activity after five reuses at pH 7.0 and 60°C. The results show that the new xylanase preparations were easily separable, thermally stable, and reusable in the synthesis of xylooligosaccharides.

Irinotecan (CPT-11), a chemotherapeutic agent used to treat several types of cancer, induces cytotoxic effects on healthy cells. The epidermal growth factor receptor (EGFR) plays a crucial role in various forms of cancer. Nimotuzumab (NmAb), a monoclonal antibody that targets the EGFR, is utilized in some countries to treat malignancies that have an overexpression of EGFR. Yet, there is a lack of literature on the potential anticancer properties of the CPT-11 and NmAb combination on in vitro human cervical cancer cells. This study investigates the apoptosis mode of the CPT-11 and NmAb combination on cervical HeLa cancer cells. The Annexin V/propidium iodide staining examination demonstrated that the combination of CPT-11 and NmAb resulted in a decrease in the number of viable cells and more potent induction of cell apoptosis than the effects of CPT-11 or NmAb alone in HeLa cells. Furthermore, the combined treatment resulted in elevated levels of reactive oxygen species (ROS) and Ca²⁺ compared to the treatment with CPT-11 or NmAb alone. Cells that were pretreated with N-acetyl-l-cysteine, a substance that scavenges ROS, and then treated with CPT-11, NmAb, or a combination of CPT-11 and NmAb exhibited higher numbers of viable cells compared to those treated with CPT-11 or NmAb alone. The combination of CPT-11 and NmAb resulted in significantly higher caspase-3, -8, and -9 activity levels than CPT-11 or NmAb alone, as measured by flow cytometer assay. The combination of CPT-11 and NmAb in HeLa cells resulted in elevated endoplasmic reticulum stress-, mitochondria-, and caspase-mediated proteins compared to treatment with CPT-11 or NmAb alone. According to these observations, NmAb enhances the effectiveness of CPT-11 in fighting cancer by stimulating cell death in the HeLa cells. Therefore, NmAb has the potential to improve the efficacy of CPT-11 as a future cervical cancer treatment in humans.

Dysfunction of the alveolar endothelial barrier plays a crucial role in the pathogenesis of septic acute lung injury (ALI). orexin B is a neuropeptide derived from orexin neurons in the lateral hypothalamus and has multiple biological functions. However, the physiological function of orexin B in sepsis is less reported. Here, we aimed to explore the protective effects of orexin B in sepsis-induced ALI and its underlying mechanisms. In this study, we established an ALI in vivo animal model in mice using cecal ligation and puncture (CLP) and an in vitro ALI model using mouse lung microvascular endothelial cells (MLMECs) induced with lipopolysaccharides (LPS). The animal experiments involved four groups: Sham, Sham+orexin B, CLP, CLP+orexin B. First, our results demonstrate that the levels of serum orexin B but not orexin A were reduced in septic mice. Correspondingly, the expression of orexin type 2 receptor (OX2R), but not orexin type 1 receptor (OX1R), was reduced in the lung tissue of septic mice. Administration of orexin B decreased the mortality in sepsis mice and improved M-CASS scores. Hematoxylin-eosin (H&E) staining assay demonstrated that administration of orexin B ameliorated histopathological lung injury. orexin B was also found to inhibit the inflammatory response in the lung tissues of septic mice by reducing the expression of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and recombinant chemokine C-X-C-motif ligand 15 (CXCL15). Additionally, the total cell count and neutrophils in bronchoalveolar lavage fluid (BALF) were reduced by orexin B. Notably, orexin B alleviated vascular endothelial permeability in mice lung tissue by increasing the expression of the tight junction protein zonula occludens-1 (ZO-1) and occludin. In vitro experiments demonstrated that orexin B prevented LPS-induced endothelial permeability in mouse lung microvascular endothelial cells (MLMECs) by upregulating the expression of ZO-1 and occludin. These effects are mediated by rho-associated coiled-coil containing protein kinase 2 (ROCK2). Based on these findings, we conclude that orexin B alleviates sepsis-induced ALI by ameliorating endothelial permeability of lung microvascular endothelial cells.

In prior research, both miRNA-125b and BLZ945 have shown potential in effectively inhibiting M2 macrophage polarization and producing antitumor effects. Nevertheless, their physicochemical characteristics present significant challenges for efficient in vivo delivery. Ionizable cationic lipid nanoparticles (LNPs), recognized for their superior biocompatibility and drug-loading capacity, serve as a novel carrier for nucleic acid-based therapeutics. In our study, we successfully encapsulated both agents within LNPs and conducted a thorough characterization. Subsequently, we investigated their potential to repolarize M2 macrophages in vitro and evaluated their in vivo distribution, biosafety, and antitumor efficacy. The findings revealed that the LNPs maintained excellent drug-loading efficiency, consistent particle size, and stable zeta potential. All formulations effectively inhibited M2 macrophage polarization in vitro. Upon administration in vivo, the LNPs not only demonstrated favorable biosafety profiles but also accumulated efficiently in tumor tissues, substantially reducing tumor burden, particularly notable in co-loaded LNPs. Our results affirm that LNPs are an effective carrier for miRNA-125b and BLZ945, highlighting this encapsulation approach as promising for the treatment of solid tumors and meriting further investigation. Practitioner points: (i) Ionizable cationic nanoparticles provide high and stable encapsulation rates to efficiently load nucleic acid polymers into the LNP, avoiding the rapid accumulation of circulating macrophages, which can lead to reduced penetration of the LNP into target tissues. Therefore, it can be used as a novel drug delivery method to benefit clinical patients. (ii) miRNA-125b LNP/BLZ945 LNP attenuated the depleting effect of BLZ945 on macrophages and significantly inhibited macrophage M2 polarization. It could be effectively distributed in tumors and showed good biosafety while exerting antitumor effects, bringing hope to clinical pancreatic tumor patients.

Metal-based nanoparticles have been extensively researched for their distinctive characteristics. Among them, zinc oxide nanoparticles have numerous applications in the field of biomedicine. The phytoextract of Ixora coccinea flowers was used in the synthesis of ZnO nanoparticles replacing the use of harmful reducing chemicals. In the current research, the carbonaceous material from biowaste of Setaria italica was used to synthesize graphene oxide (GO) by Improved Hummer's method. The synthesized GO was converted to reduced GO via green nanotechnology using phytoextract of Prosopis juliflora. The synthesis of reduced Graphene Oxide - Zinc Oxide Nanocomposite (rGO)-ZnO nanocomposite involves a simple, economical one-step magnetic stirring method. UV-visible spectroscopy was used to characterize the synthesized materials, with the maximal absorbance range for ZInc Oxide (ZnO) being 384 nm and for rGO-ZnO composite at 243 and 366 nm, respectively. The x-ray diffraction (XRD) revealed 2θ peaks for ZnO at 31.54°, 34.22°, and 36.08°. For reduced Graphene Oxide (rGO) in rGO-ZnO composite, the XRD revealed 2θ peaks at 21.25°, 21.56°, 23.14°, and for ZnO at 31.74°, 33.24°, 34.29°, 36.23°. The FT-IR demonstrated the vibrational modes of functional groups: -OH stretching, symmetric and antisymmetric -CH₂ stretching, C = C stretching, and C-O stretching. The elemental composition of samples has been analyzed using Energy Dispersive x-ray spectroscop (EDX), and the high percentage of zinc in the composite shows a good loading rate of ZnO on the rGO's surface. By morphological investigation, monolayer sheet structures of rGO loaded with clusters of ZnO are clearly demonstrated. Positive results from therapeutic assays and biocompatibility were found with reduced hemolysis and good anticoagulation abilities proved with statistical approach. Our research is distinctive because a realistic formulation of an rGO-ZnO skin care cream with enhanced therapeutic properties, such as effective stability, spreadability, and significant moisture retention, can be recommended.

Enhancing the biocompatibility of biomaterials is a critical aspect of tissue engineering and regenerative medicine. Advances in 3D bioprinting technology, blending natural and synthetic materials for the production of bioink, offer new opportunities to develop highly biocompatible materials that can closely mimic the native tissue environment. In this study, we used pericardial fluid structure (PFS)-based material together with alginate to mimic the extracellular matrix (ECM) and produce a bioink material. Thus, blended alginate with PFS material and MC3T3-E1 pre-osteoblast cell-laden hydrogels characterized by comparing each other, especially alginate hydrogels, and evaluated in terms of biocompatibility for tissue engineering applications. According to the rheological analysis results, all hydrogel groups A, A-PFS (150 mg), and A-PFS (1:1) had viscoelastic properties. Mechanical tests showed that the A-PFS (1:1) hydrogel had the most strength properties. Additionally, the viscosity values of the hydrogel solutions were in an applicable range for use in 3D bioprinters. It was also found out that PFS increased the biocompatibility of alginate-based bioink, in terms of cell proliferation and differentiation. Overall, these findings suggest that alginate and pericardial fluid-based materials can be successfully used for bioink production. The resulting hydrogels exhibit viscoelastic properties, appropriate viscosity for 3D bioprinting, and support cell viability, proliferation, and osteogenic differentiation. This research has the potential not only to produce bioink but also to produce injectable hydrogels and drug delivery systems, which can become biocompatible materials that can be used for tissue engineering and regenerative medicine applications.

Checkpoint inhibitors are widely recognized immunotherapeutic drugs, known for their effectiveness in treating various cancers. Atezolizumab, targeting the immune checkpoint programmed death-ligand 1, is successfully used to treat several types of cancers. Atezolizumab is a potential biosimilar candidate due to its huge success in the clinic but there is no literature on its production process in mammalian cells. In this study, we generated a monoclonal cell line derived from recombinant Chinese hamster ovary DG44 cells to produce atezolizumab. The selected single clone was employed for media screening and process development. Following production in a 7-L bioreactor, atezolizumab was purified using a three-step chromatographic method. Finally, the purified atezolizumab was characterized and compared with commercial atezolizumab (Tecentriq) through several chromatographic and kinetics analyses.

Triple-negative breast cancer (TNBC), known for its hostile nature and limited treatment modalities, has spurred researchers to explore novel approaches for enhancing clinical outcomes. Here, the study aimed to analyze transcriptomics data to identify immune-related hub genes associated with TNBC that might serve as prognostic biomarkers. Initially, we determined genes that were differentially expressed between TNBC and normal tissues by integrating microarray and RNA sequencing data. Then, through protein-protein interaction and module analysis, we identified five putative hub genes: AURKA, CCNB1, CDCA8, GAPDH, and TOP2A. Subsequently, gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that the hub genes were primarily involved in the progesterone-mediated oocyte maturation signaling pathway and oocyte meiosis. Additionally, we observed that these five hub genes were significantly elevated at both protein and mRNA levels in TNBC tissues and contributed to worse survival. Furthermore, the expression of these hub genes exhibited a strong positive association with immune-invading cells such as CD8 T cells, CD4 T cells, and dendritic cells. The analysis of the regulatory network revealed three transcription factors (YBX-1, E2F1, and E2F3) and three posttranscriptional regulators (hsa-mir-25-3p, hsa-mir-92a-3p, and hsa-let-7b-5p) of hub genes. Finally, we explored potential drug candidates for the hub genes using Drug-Gene Interaction Database and discovered that there are no FDA-approved drugs for CCNB1 and CDCA8, highlighting a promising area for future research. Taken together, our results will be of immense importance in addressing the intricacies of TNBC.