BMC Biotechnology最新文献

Background: The Arctic environment, characterized by extreme conditions, hosts a largely untapped reservoir of fungal communities that have adapted to these harsh conditions by producing specialized bioactive secondary metabolites. Among these, Acremonium species also remain underexplored, despite their potential to produce structurally diverse and biologically active secondary metabolites. This is largely due to difficulties in sampling in remote Arctic regions and limited research focus on fungi from such environments. This study aimed to use an integrated workflow combining metabolomics, chemometrics, and bioactivity screening to prioritize Acremonium strains for the identification of bioactive secondary metabolites. We applied this workflow to investigate six Acremonium strains associated with driftwood from the Arctic: A. ellipsoideum (F1, F2), A. synnematoferum (F3, F4, F5), and A. multiramosum (F6), aiming to identify cytotoxic secondary metabolites.

Results: The integrated metabolomics and chemometrics approach revealed unique chemical fingerprints for A. ellipsoideum (F1) and A. synnematoferum (F5) among the six strains. By further combining bioactivity screening results, strain F5 was prioritized for further detailed study. Five compounds were isolated from F5 and structurally elucidated as cyclic depsipeptides: destruxin-A4 chlorohydrin (1), trichomide D (2), destruxin-A5 (3), homodestruxin (4), and homodestcardin (5). All compounds exhibited cytotoxic effects against the tested cell lines but did not exhibit activity in the targeted bioassays against the kinase FLT3 or the phosphatase PTP1B. Cytotoxic destruxin-type compounds likely play a significant ecological role, as they have been reported to exhibit entomopathogenic, nematocidal, and phytotoxic effects.

Conclusion: The identification of five known cytotoxic destruxin-type depsipeptides from A. synnematoferum as a new source expands the chemical diversity known for this genus and underscores their potential for the development of cytotoxic agents. These findings highlight not only the value of Arctic fungi as a reservoir of bioactive compounds but also the necessity of deciphering the ecological roles of cytotoxic metabolites produced by these organisms as they adapt to extreme environments. Furthermore, this study highlights the effectiveness of multi-informative-driven strain prioritization in uncovering bioactive metabolites from new fungal sources, emphasizing the significance of exploring Arctic fungal diversity for its potential to enhance chemical diversity, contribute to drug development, and broaden our understanding of ecological roles.

Active pharmaceutical ingredients (APIs) have poor efficacy and low bloodstream and target site concentrations due to their limited water solubility. Pregabalin is essential product to control pain and inflammation. We have synthesized novel pregabalin derivative called gabsali ((S, E)-3-(((2-hydroxybenzylidene)amino)methyl)-5-methylhexanoic acid. We have synthesized it by using salicylaldehyde to functionalize pregabalin (PG). The research was divided into two parts. The first part of the process was making a new kind of pregabalin called gabsali (GS). Second, MSNs were created using the sol-gel technique and then the synthesized GS was added to them. This study introduces a unique drug delivery method that improves bioavailability, stability, and anti-inflammatory activity by incorporating chemically modified pregabalin (GS) into engineered MSNs. This strategy has not been documented in the literature before. Zeta sizer, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), BET analysis, and high-performance liquid chromatography (HPLC) were some of the modern tools used to determine particle size, shape, drug release, and compatibility. Under all tested conditions, the inclusion compound showed a considerable improvement in dissolving rate relative to the crystalline medicine in in vitro dissolving assays. The produced MSNs had a particle size of about 500 nm and were nano-sized, spherical. The FTIR spectroscopy was used to examine the chemical GS. The medicine was successfully added into blank MSNs, resulting in a decrease in both their specific surface area (602.5 ± 0.7 m²/g) and pore width (5.9 nm). In order to identify the potential benefits of GS, this study evaluated a preclinical model of inflammation that was created using ovalbumin (OVA). In a comparison to pregabalin, its anti-inflammatory activity was tested at doses of 50, 75, and 100 mg/kg. Using reverse transcription-polymerase chain reaction (RT-PCR), the levels of inflammatory mediators (cytokines IL-2 and IL-6) in the blood were measured and discovered to be significantly reduced. Cell viability was evaluated using the MTT assay, which showed a higher IC50 compared to pregabalin, the unmodified medicine. According to in vivo pharmacokinetic studies, MSN-GS significantly increased bioavailability compared to the pure drug. Using MSN-GS, the aforementioned results clearly show biocompatibility, improved in-vivo bioavailability, and satisfactory in-vitro performance.

Background: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based gene editing has become a promising approach for enhancing traits in aquaculture species. Nevertheless, traditional CRISPR-Cas systems encounter challenges, including significant off-target effects and strict protospacer adjacent motif (PAM) requirements, which constrain their use in crustaceans such as Penaeus monodon. To address these limitations, this research has developed PmMAD7, a codon-optimized CRISPR system specifically designed for P. monodon, which incorporates nuclear localization signals to improve editing efficiency and precision.

Results: This research successfully synthesized and delivered PmMAD7 mRNA and crRNAs targeting the ECH1 and AQP4 genes into the hemocytes of P. monodon. Quantitative PCR analysis demonstrated that PmMAD7 achieved significant gene silencing, reducing the expression levels of ECH1 and AQP4 by 81.5% and 78.33%, respectively. Next-generation sequencing confirmed targeted insertions and deletions at the gene loci, with knockout efficiencies of 14.81% for ECH1 and 20.57% for AQP4, which were significantly higher than those obtained with LbCas12a (7.14% and 12.43%, respectively). Furthermore, functional analysis indicated that ECH1 knockout resulted in increased cell volume and mortality, while AQP4 knockout led to decreased cell volume and reduced viability. These specific results highlight the first successful demonstration of MAD7-based genome editing in shrimp. The broader PAM compatibility and enhanced editing efficiency of PmMAD7 provide a versatile platform for gene editing in shrimp.

Conclusion: PmMAD7 constitutes an enhanced CRISPR editing tool specifically designed for P. monodon, exhibiting superior precision, expanded PAM compatibility, and enhanced editing efficacy relative to conventional Cas12a systems. These results lay the groundwork for the advancement of gene editing applications in crustaceans and contribute to sustainable genetic improvements in aquaculture.

Background: Early-stage Luminal A breast cancer generally has a favorable prognosis, yet some patients experience recurrence, presenting a challenge in understanding the underlying genetic factors. This study aimed to identify genetic mutations associated with recurrence in early-stage Luminal A breast cancer patients through whole-exome sequencing (WES).

Methods: We collected formalin-fixed paraffin-embedded samples from 34 patients and divided them into two groups: 17 patients with recurrence within five years post-surgery (recurrence group) and 17 patients with no recurrence for over five years (control group). The extracted DNA went through library preparation and was subjected to WES. Sequencing data went through quality control, alignment, and mutation identification. Functional enrichment analyses were conducted to explore the biological implications of the mutations.

Results: We generated on average ~ 11 Gb raw sequencing data for each sample and identified 7,066 nonsynonymous mutations. The recurrence group exhibited a higher mutation rate (11.48 mutations/Mb) compared with the control group (9.18 mutations/Mb, p < 0.05). A significant negative correlation was observed between disease-free survival time and the number of mutations (p < 0.05). Eight genes (MICALCL, G6PD, OR8U1, PCLO, OR8U8, ZCCHC18, CPED1, HMCN1) were significantly associated with early recurrence (p < 0.05). Functional enrichment analyses revealed that these genes were involved in pathways like mismatch repair and immune response.

Conclusions: This study identified specific genetic mutations linked to early recurrence in Luminal A breast cancer, highlighting potential biomarkers for predicting patient outcomes and personalizing cancer treatment. Our study also showed that state-of-the-art WES can extract biologically and clinically meaningful mutation signatures from routinely stored FFPE tissues, unlocking archived specimens for large-scale biomarker discovery.

The present study focused on the antibacterial activity, mechanism and application of Pseudoduganella armeniaca ZMN - 3 extracellular polysaccharide nano silver. Extracellular polysaccharide nano silver was prepared by chemical synthesis and characterized by UV - Vis spectroscopy, Fourier transform infrared spectroscopy, X - ray diffraction, X - ray photoelectron spectroscopy, scanning electron microscopy and transmission electron microscopy. The antibacterial effects of extracellular polysaccharide nano silver on two common pathogens were investigated by agar well diffusion method and broth dilution method. The results showed that extracellular polysaccharide nano silver had significant inhibitory effects on Escherichia coli and Staphylococcus aureus. In addition, by SEM and TEM observation, the conductivity, the leakage of nucleic acids and proteins, the release of reducing sugars, the ATP content, and the alkaline phosphatase level were also measured. The study results indicated that extracellular polysaccharide nano silver could cause the leakage of large biomolecules such as nucleic acids and proteins, reduce the release of sugars, increase ATP content, and lead to the leakage of alkaline phosphatase, ultimately resulting in bacterial death. Finally, the application of extracellular polysaccharide nano silver in antibacterial coating was studied. Antibacterial coating achieved an impressive inhibition rate of 99% against both Escherichia coli and Staphylococcus aureus and could effectively resist bacterial adhesion within 7 days. This study provides a theoretical basis for the potential application of Pseudoduganella armeniaca ZMN - 3 extracellular polysaccharide nano silver in the antibacterial field.

Reactive oxygen species (ROS) in the tumor microenvironment (TME) are key factors involved in inhibiting tumor cell proliferation. This study developed a hollow mesoporous silica-Prussian blue (HMSNs-PB) nanozyme delivery platform loaded with metformin (HPB@MET), which enhances ROS levels within tumor cells through peroxidase-like (POD) activity. Moreover, in the acidic microenvironment of tumors, HPB@MET disintegrates, and the released MET causes mitochondrial dysfunction and increased electron leakage, ultimately promoting the abnormal accumulation of ROS in the TME. In vitro experiments demonstrate that HPB@MET achieves peak cellular uptake at 72 h and significantly inhibits tumor cell proliferation through its exceptional ROS-generating capacity. In the K7M2 osteosarcoma model (initial tumor volume 80 mm³), HPB@MET increased the tumor inhibition rate from 55% with HPB to 76.5%, with median survival extended to 57 days, indicating that HPB@MET synergistically suppresses tumor cell proliferation via POD activity and mitochondrial dysfunction, effectively suppressing osteosarcoma progression. These results indicate that this nanozyme-drug delivery platform can synergistically promote ROS generation in the TME, effectively inhibiting OS and providing a key technological approach for the efficient treatment of this disease.