Hasil untuk "Biotechnology"

Yi Shuai, Bo Chen, Tao Jiang et al.

Abstract Background Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a severe, therapy-refractory condition driven by ferroptotic disruption of jawbone-derived mesenchymal stem cells (MSCs) biology. We dissect this mechanism to validate ferroptosis as a therapeutic target. Methods We first demonstrated that miR-145-3p is mechanistically coupled to ferroptosis and osteogenesis in BRONJ model by gain- and loss-of-function studies. To evaluate therapeutic efficacy under pathologically relevant conditions, we designed three models: (i) Local BRONJ repair model: miR-145-3p-enriched exosomes were encapsulated in an injectable hydrogel scaffold and grafted into necrotic alveolar bone to assess direct BRONJ resolution. After local BRONJ treatment, endogenous MSCs were re-isolated, and both cellular and exosomal miR-145-3p levels were quantified. (ii) Osteoporosis treatment model: The reprogrammed MSCs derived from treated BRONJ rats were then administered intravenously to osteoporotic littermates to evaluate whether these MSCs retain systemic osteogenic competence. (iii) Critical-sized calvarial defect repair model: To further dissect the intrinsic osteogenic capacity, reprogrammed MSCs derived from treated BRONJ rats were fabricated into cell-sheet/HA-TCP “sandwich” constructs and transplanted into calvarial defects. Results BRONJ markedly compromised MSCs viability while elevating hallmarks of ferroptosis that were reversed by the ferroptosis inhibitor. Concomitantly, osteogenic capacity declined, as shown by reduced ALP activity, mineralized nodules, new bone formation and expression of RUNX2 and OCN. Mechanistically, we identified a miR-145-3p/IREB2 regulatory circuit that governs ferroptosis in BRONJ-derived MSCs; exosome-mediated delivery of miR-145-3p reinstated this axis, thereby reactivating the MSCs osteogenesis and driving in situ bone regeneration. Critically, after local BRONJ therapy, the reprogrammed MSCs were (i) infused via tail vein into osteoporotic rats, significantly elevating bone mass, and (ii) engineered into cell-sheet/HA-TCP “sandwich” constructs that achieved robust repair of critical-sized calvarial defects. These data establish exosomal miR-145-3p as a therapeutic that mitigates ferroptosis and restores the osteogenic competence of jawbone-derived MSCs for bone regeneration. Conclusion Collectively, our findings establish the miR-145-3p/IREB2/ferroptosis axis as an important regulator of BRONJ pathology and demonstrate that exosomal delivery of miR-145-3p not only ameliorates localized BRONJ but also substantially reinstates the systemic osteogenic potential of jawbone-derived MSCs, offering a pre-clinical promising strategy to combat both BRONJ and associated bone loss disorders. Graphical Abstract

Kecan Chen, Runcheng Zhou, Wenjing Zhang et al.

Abstract Streptococcus mutans mediates enamel demineralization through acid production via glycolysis, while Streptococcus salivarius, as a commensal bacterium, promotes caries progression by enhancing biofilm formation. Their synergistic interaction amplifies cariogenicity. Therefore, developing strategies to inhibit both bacterial species is imperative. This study investigated the extraction and characterization of a polysaccharide from mangosteen scarfskin (MSP) and its antimicrobial potential against cariogenic bacteria. Using ultrasonic-assisted extraction, MSP was obtained with a yield of (9.93 ± 0.5696)%, presenting light brown coloration. Antimicrobial assays demonstrated strong anti-efficacy activity against Streptococcus mutans and Streptococcus salivarius, showing a MIC of 1 mg/mL and significant bactericidal effects at 1×MIC and 2×MIC concentrations. Biofilm metabolism analysis showed that MSP caused dose-dependent suppression of bacterial metabolism, while its inhibitory effect on EPS production increased proportionally with concentration. Molecular docking identified specific hydrogen-bond interactions between arabinose (the primary component of MSP) and key residues (THR-315, SER-10, and SER-247) of glucosyltransferase-C (GTF-C), while molecular dynamics simulations demonstrated that arabinose disrupted the structural stability of GTF-C. These findings collectively suggest MSP’s promising application as a novel food additive for caries prevention through oral streptococcal control. Graphical abstract

Guohua Bao, Xiao Xu, Jing Yang et al.

Leaf senescence is an important agronomic trait that significantly influences the quality and yield of soybeans. v-Myb avian myeloblastosis viral oncogene homolog (MYB) transcription factors are considered crucial regulators governing leaf senescence, which can be utilized to improve agronomic traits in crops. However, our knowledge regarding the functional roles of soybean MYBs in leaf senescence is extremely limited. In this study, GmMYBLJ, a CCA1-like MYB, was identified and functionally characterized with respect to leaf senescence. The GmMYBLJ protein is localized in the nucleus, and a high accumulation of its transcripts was observed in nodules and embryos. Notably, GmMYBLJ was highly expressed in soybean senescent leaves and was transcriptionally induced by dark or NaCl treatment, as confirmed by histochemical GUS staining analysis. Ectopic overexpression of GmMYBLJ in Arabidopsis not only led to earlier leaf senescence, reduced chlorophyll content, and increased MDA accumulation but also promoted the expression of several WRKY family transcription factors and senescence-associated genes, such as SAG12 and ORE1. Further investigation showed that overexpression of GmMYBLJ accelerated Arabidopsis leaf senescence under darkness and in response to Pst DC3000 infection. Moreover, transgenic soybean plants overexpressing GmMYBLJ grew faster and exhibited accelerated senescence under salt stress. DAB staining analysis showed that GmMYBLJ induced ROS accumulation in soybean hairy roots and Arabidopsis leaves. Collectively, our results provided useful information into the functional roles of GmMYBLJ in both age-dependent and stress-induced senescence.

Rishi Ram, Bhawna, Sanjeev Kumar et al.

IntroductionPesticides such as isoproturon are widely employed and represent a considerable environmental concern. The development of sustainable and efficient degrading techniques is crucial. Photocatalytic degradation employing semiconductor materials is a compelling solution. This study examines the synergistic advantages of heterojunction formation by synthesizing, characterizing, and improving the photocatalytic efficacy of Ag3PO4/SnO2 nanocomposites for the degradation of isoproturon.MethodsThe Ag3PO4/SnO2 nanocomposite was characterised using powder X-ray diffraction (PXRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Ultraviolet-Diffuse Reflectance Spectroscopy (UV-DRS) and X-ray Photoelectron Spectroscopy (XPS). The effective synthesis of the Ag3PO4/SnO2 heterojunction was confirmed by characterization data from various techniques (PXRD, FTIR, SEM, UV-DRS, XPS).Results and DiscussionElemental mapping confirmed uniform distribution of O, P, Ag, and Sn. High-resolution mass spectrometry (HRMS) was employed to analyse degradation products. The Ag3PO4/SnO2 nanocomposite exhibited improved photocatalytic degradation of isoproturon compared to its precursors. In contrast to 25% for pure SnO2 and 41% for Ag3PO4, over 97% degradation was achieved using Ag3PO4/SnO2 nanocomposite within 120 min of light irradiation under identical conditions. The synergistic effects of heterojunction formation significantly enhanced isoproturon degradation using the Ag3PO4/SnO2 nanocomposite. The heterojunction reduces electron-hole recombination rate and enhances photogenerated charge carriers for degradation via effective charge separation. The improved photocatalytic activity is ascribed to the increased surface area of the nanocomposite. The analysis of HRMS data revealed the degradation products. The findings demonstrate the efficacy of Ag3PO4/SnO2 nanocomposites as photocatalysts for environmental remediation, namely in the breakdown of pesticides.

Xinyue Gao, Feichang Liu, Bo Zhang et al.

Abstract Cell division cycle 25 (CDC25) phosphatases serve as crucial regulators of cell cycle phase transitions and essential components of the checkpoint machinery involved in DNA damage response. Emerging evidence indicates the oncogenic potential of CDC25 family members across various cancers. However, comprehensive insights into the expression pattern and function of the CDC25 family in diverse cancers remain unexplored. In our study, we investigated CDC25 family using multiple databases, including gene expression levels, molecular signatures, diagnosis value, and prognostic value in pan-cancer. Furthermore, we focused on melanoma and systematically explored CDC25A expression and its clinical correlations. As a result, the expression of CDC25 family members is significantly abnormal in most cancers, correlating with poorer prognosis. CDC25 family members are differently regulated by DNA methylation and genetic alterations across various cancers. In addition, CDC25 family plays a critical role in the malignant progression of melanoma. Functional investigation reveals that CDC25A inhibition suppresses the proliferation of melanoma cells and sensitizes melanoma cells to chemotherapy and NK cell therapy. In conclusion, our study suggests that CDC25 family may serve as a significant biomarker for diagnosis and prognosis across multiple cancers, with CDC25A as a promising therapeutic target for melanoma.

Nan Zheng, Yongchao Cai, Zehua Zhang et al.

Abstract The pursuit of obtaining enzymes with high activity and stability remains a grail in enzyme evolution due to the stability-activity trade-off. Here, we develop an isothermal compressibility-assisted dynamic squeezing index perturbation engineering (iCASE) strategy to construct hierarchical modular networks for enzymes of varying complexity. Molecular mechanism analysis elucidates that the peak of adaptive evolution is reached through a structural response mechanism among variants. Furthermore, this dynamic response predictive model using structure-based supervised machine learning is established to predict enzyme function and fitness, demonstrating robust performance across different datasets and reliable prediction for epistasis. The universality of the iCASE strategy is validated by four sorts of enzymes with different structures and catalytic types. This machine learning-based iCASE strategy provides guidance for future research on the fitness evolution of enzymes.

Han Na Suh, Sung-Hoon Yoon, Yoo Min Park et al.

Elena Laura Mazzoldi, Giulia Gaudenzi, Paola Serena Ginestra et al.

IntroductionTissue engineering has advanced significantly in recent years, owing primarily to additive manufacturing technology and the combination of biomaterials and cells known as 3D cell printing or Bioprinting. Nonetheless, various obstacles remain developing adequate 3D printed structures for biomedical applications, including bioinks optimization to meet biocompatibility and printability standards. Hydrogels are among the most intriguing bioinks because they mimic the natural extracellular matrix found in connective tissues and can create a highly hydrated environment that promotes cell attachment and proliferation; however, their mechanical properties are weak and difficult to control, making it difficult to print a proper 3D structure.MethodsIn this research, hydrogels based on Alginate and Gelatin are tested to evaluate the metabolic activity, going beyond the qualitative evaluation of cell viability. The easy-to-make hydrogel has been chosen due to the osmotic requirements of the cells for their metabolism, and the possibility to combine temperature and chemical crosslinking. Different compositions (%w/v) are tested (8% gel-7% alg, 4% gel-4% alg, 4% gel-2% alg), in order to obtain a 3D structure up to 10.3 ± 1.4 mm.ResultsThe goal of this paper is to validate the obtained cell-laden 3D structures in terms of cell metabolic activity up to 7 days, further highlighting the difference between printed and not printed cell-laden hydrogels. To this end, MS5 cells viability is determined by implementing the live/dead staining with the analysis of the cellular metabolic activity through ATP assay, enhancing the evaluation of the actual cells activity over cells number.DiscussionThe results of the two tests are not always comparable, indicating that they are not interchangeable but provide complementary pieces of information.

Mirza Abrar, Tanveer Ahmad, Shahid Iqbal et al.

Abstract The genus amaranth is comprised of 60–70 species. This climate hardy plant is grown as a vegetable as well as cereal crop in many areas of the world. Current study multivariate analysis was used to characterize the exotic vegetable amaranth genotypes. The experiment was conducted in the research area of Muhammad Nawaz Shareef University of Agriculture, Multan under RCBD design with three replications of 50 exotic amaranth genotype. 22 different variables were investigated for the principal component and cluster analysis using different software packages in R studio. According to the results of principal component analysis the first five PCs had eigenvalues greater than one and they explained the 84.59% diversity in exotic vegetable amaranth accessions. First two principal components explained 57.31% of variation. Traits that loaded highly for first PC include moisture (-0.892), carbohydrates (0.938), energy (0.920), stomatal conductance (0.803), net photosynthesis (0.795) and dry weight (0.770). The traits which impacted the second PC highly include protein (0.692), fat (-0.571), nitrogen (0.692), calcium (0.545), phosphorus (0.908), zinc (0.960), iron (0.826) and potassium (0.872). For future breeding program the selection based on these results will be effective for crop enhancement drive in exotic vegetable amaranth genotypes. Hierarchical clustering was done and dendrogram split the 50 genotypes into 5 groups using silhouette method. The pairwise clustering revealed that intra cluster distance is smaller than inter cluster distance. The maximum inter cluster distance was observed between cluster I and II (9.88); between cluster I and III (9.47); between cluster II and IV (9.27). Clinical trial number Clinical trial number not applicable.

Salam S Alsharari, Muneefah A. Alenezi, Mona S. Al Tami et al.

A critical milestone in nano-biotechnology is establishing reliable and ecological friendly methods for fabricating metal oxide NPs. Because of their great biodegradable, electrical, mechanical, and optical qualities, zirconia NPs (ZrO2NPs) attract much interest among all zirconia NPs (ZrO2NPs). Zirconium oxide (ZrO2) has piqued the interest of researchers throughout the world, particularly since the development of methods for the manufacture of nano-sized particles. An extensive study into the creation of nanoparticles utilizing various synthetic techniques and their potential uses has been stimulated by their high luminous efficiency, wide bandgap, and high exciton binding energy. Zirconium dioxide nanoparticles may be used as antimicrobial and anticancer agents in food packaging. In response to the growing interest in nano ZrO2, researchers invented and developed methods for synthesizing nanoparticles. ZrO2 nanocomposites with various morphologies have recently been created using biological (green chemistry) methods. Microbes and plants both contribute to the production of zirconia in the laboratory. Capping and stabilizing agents are provided by the biomolecules found in plant extracts, whereas microorganisms provide enzymes as capping and stabilizing agents (intracellular or extracellular). It is possible to analyze the nanoparticles produced using a variety of analytical approaches, including ultraviolet-visible spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FT-IR). When applied to bacteria (both Gram-positive and Gram-negative) and fungi, ZrO2NPs show promising antibacterial capabilities. Normal and malignant cells are sensitive to ZrO2 nanoparticles, which can be explained by the generation of reactive oxygen (ROS). This work discusses and describes many ways of producing ZrO2 nanoparticles, their properties, and various application possibilities.

Masaru Murata, Md Arafat Kabir, Yukito Hirose et al.

The aim of this clinical case study was to observe biopsy tissues at 5 months after an autograft of a partially demineralized dentin/cementum matrix (pDDM) into a tooth-extracted socket exhibiting healing failure. A 66-year-old female presented with healing failure in the cavity for 2 months after the extraction (#36). Initial X-ray photos showed a clear remainder of lamina dura (#36), a residual root (#37), and a horizontal impaction (#38). The vital tooth (#38) was selected for pDDM. The third molar crushed by electric mill was decalcified in 1.0 L of 2.0% HNO₃ for 20 min and rinsed in cold distilled water. The pDDM granules (size: 0.5–2.0 mm) were grafted immediately into the treated socket. X-ray views just after pDDM graft showed radio-opaque granules. At 5 months after pDDM graft, the surface of regenerated bone was harmonized with the mandibular line, and bone-like radio-opacity was found in the graft region. The biopsy tissue (diameter: 3.0 mm) at 5 months after pDDM graft showed that mature bone was interconnected with the remaining pDDM. The novel histological evidence highlighted that newly formed bone was connected directly with both dentin-area and cementum-area matrix of pDDM. We concluded that pDDM contributed to the regeneration of bone in the unhealed socket, and this regeneration prepared the socket for implant placement. Autogenous pDDM could be immediately recycled as an innovative biomaterial for local bone regeneration.

Yi Li, Yue Zang, Tianda Fan et al.

Up to 20% of patients treated with anti-PD-1/PD-L1 inhibitors suffered from thyroid dysfunctions, yet the mediators associated with their occurrence remain unclear. The increasing coincidence of papillary thyroid carcinoma (PTC) with Hashimoto thyroiditis (HT) and the high vulnerability of thyroid to immunotherapy motivated us to discover the similarities and their underlying transcriptomic basis. Clinical characteristics analysis of 468 PTC patients from two independent cohorts and meta-analysis of 22,155 PTC patients unveiled a strong negative association between HT and recurrence in PTC patients. Transcriptome analysis of both cohorts showed PTC patients with HT were enriched in macrophages, CD8+ and CD4+ cytotoxic T cells, which was further validated by single-cell transcriptome analysis of 17,438 cells from PTC patients, and CD8+ T cells were correlated with disease-free survival of PTC patients. In both cohorts and single-cell dataset, elevated expression of PD-1-related genes was observed in the HT group, and CD3D appeared to be a target for enhancing the activation of CD8+ T cells. Correlation analysis of 3,318 thyroid adverse events from 39,123 patients across 24 tumor types and molecular signatures demonstrated similar signatures associated with autoimmune thyroiditis in PTC and thyroid immune-related adverse events (irAEs), and several multi-omics signatures, including signatures of CD8A and CD8+ T cells, showed positive associations with the odds ratio of thyroid irAEs. Our results unveil shared molecular signatures underlying thyroid dysfunction between patients receiving immunotherapies and PTC patients suffering from HT, which may shed light on managing the adverse events during cancer immunotherapy.

Allali Aimad, Mohammed Bourhia, Hadin Hana et al.

Callosobruchus maculatus (Fab.) (C. maculatus) is one of the major pests of legume seeds in storage causing significant damage, leading to food insecurity and low income for farmers. This work was planned to develop eco-friendly agents from essential oils of Artemisia herba alba Asso. (AEO), Maticaria Recutita L. (MEO), and Dittrichia Viscosa L. (DEO) to control C. maculatus. To achieve this goal, essential oils (EOs) were extracted by hydro-distillation using Clevenger apparatus before being characterized by GC-MS. EOs were used for testing purposes using three different tests, namely, inhalation toxicity, contact toxicity, and repellency tests. GC-MS analysis of EOs showed the presence of 16 potentially active compounds in AEO and 38 in MEO, whilst 15 compounds were identified in DEO. AEO was higher in thujone (57.6%) and chrysanthenone (11.8%). Santolina alcohol (40.7%) and germacrene D (8.9%) were the major compounds identified in MEO, whereas isocostic acid (72.3%) was the chief compound of DEO. The obtained findings showed that the studied EOs showed considerable insecticidal activity against C. maculatus with a lethal dose (LC50) of 3.78, 8.86, and 14.34 μL/1 liter of air by AEO, MEO, and DEO, respectively. At 1 μL/1 liter of air, the oviposition reduction rate was 90.02%, 70.65%, and 48.23% by AEO, MEO, and DEO, respectively, whereas the emergence reduction rate was 87.32%, 60.08%, and 32.24% by AEO, MEO, and DEO, respectively. With increasing doses up to 20 μL/L, the reduction of individual emergence reached 98.8% by AEO of 24 h after treatment. AEO, MEO, and DEO showed significant repellent effects against adults of C. maculatus with repulsion percentages of 60.83%, 50.83%, and 72.5%, respectively. The outcome of this work suggests that the essential oils of the studied plants, particularly Artemisia herba alba Asso. oils, can constitute a natural and environmentally friendly alternative to develop new bioinsecticides for the control of C. maculatus.

P. Haaland

Chunyan Liu, Mengzhen Xu, Luocheng Yan et al.

Abstract Background Honeysuckle is a time‐honored herb with anticancer activity in traditional Chinese medicine. Recently, accumulating reports are suggesting that the microRNAs in this medicinal plant not only play a physiological role in their original system, but also can be transmitted to another species as potential therapeutic components. In the numerous bioactive investigations, the anti-tumor effects of these microRNAs in the magical herb are rarely studied, especially the special miR2911, a honeysuckle-encoded atypical microRNA, with high stability during the boiling process and unique biological activity to target TGF-β1 mRNA. Methods Luciferase assay was conducted to test the ability of miR2911 to target TGF-β1 mRNA. ELISA was performed to determine the expression level of TGF-β1 of mouse colorectal adenocarcinoma CT26 cells when treated with miR2911 and tumor tissue in Sidt1 +/+ and Sidt1 −/− mice. qRT-PCR was performed to examine the level of expression of miR2911. Tumor-bearing wild and nude mice were employed to evaluate the anti-tumor effect of honeysuckle and miR2911 in vivo. Tumor tissue necrosis was observed by H&E staining. Besides, the infiltration of T lymphocytes across solid tumors was tested by immunostaining staining. Results Our results showed that honeysuckle slowed the development of colon cancer down. Further research showed that miR2911 could bind strongly to TGF-β1 mRNA and down-regulate the expression of TGF-β1 and had a high stability under boiling and acid condition. Moreover, SIDT1 mediated dietary miR2911 inter-species absorption. And we found that miR2911 had a similar anticancer effect as honeysuckle. Mechanistically, miR2911 reversed the tumor-promoting effect of TGF-β1 by an increase of T lymphocytes infiltration, resulting in slowing the colon cancer process in immunocompetent mice. Consistent with this inference, the anti-tumor effect of miR2911 was revealed to be abolished in T cell immune deficiency mice. Conclusion Taken together, honeysuckle-derived miR2911 showed an anti-tumor effect in colon cancer through targeting TGF-β1 mRNA. The down-regulation of TGF-β1 promoted T lymphocytes infiltration, and accordingly impeded the colon tumor development.

P. Velho

J. Benemann

E. Bayer, E. Morag, R. Lamed

N. Jenkins, R. Parekh, D. James

V. Shiva