BMC BiotechnologyPub Date : 2025-05-24DOI: 10.1186/s12896-025-00981-7
Teslim Yimam, Mereme Abide, Solomon Benor, Demisachew Guadie
{"title":"Protocol optimization for callus induction and shoot regeneration of Ethiopian rice varieties (Oryza sativa L.).","authors":"Teslim Yimam, Mereme Abide, Solomon Benor, Demisachew Guadie","doi":"10.1186/s12896-025-00981-7","DOIUrl":"10.1186/s12896-025-00981-7","url":null,"abstract":"<p><p>Rice is the primary food source for the majority of the global population. In African countries, rice is considered a critical cereal crop essential for food security. Rice consumption in Ethiopia has been increasing from year to year, and the demand for rice food is not fulfilled by domestic production, the Ethiopian government imports rice products from Asia. Therefore increasing the production of rice in terms of quantity and quality through modern technology is a recent priority and strategy in the country. This study was, therefore, designed to optimize the protocol for callus induction and shoot regeneration of two Ethiopian rice (X-Jigna and Shaga) varieties. Three media types (LS, N6, and MS) supplemented with different concentrations of 2, 4-D were used for callus induction. The maximum callus induction was observed on MS-media supplemented with 2.5 mg/L for X-Jigna and Shaga varieties, respectively. The lowest callus induction frequencies were recorded on N6-media supplemented with 1.5 mg/L 2, 4-D for both varieties. The highest shoot regeneration for X-Jigna (80.67%) and Shaga (72%) was recorded on MS-media containing a combination of kinetin (2 mg/L) and Naphtalin Acetic Acid (0.2 mg/L). The survival rate of acclimatized plantlets was 100% and 66.6% for X-Jigna and Shaga, respectively. In conclusion, this study is distinctive as, to our knowledge, no other research has addressed the optimization of protocols for the Ethiopian rice varieties, serving as a foundational element for future investigations into rice transformation and breeding in Ethiopia.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"43"},"PeriodicalIF":3.5,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103809/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-22DOI: 10.1186/s12896-025-00964-8
Heba A H Zaghloul, Zhengkun Xiao, Hengrui Hu, Guo-Hua Huang
{"title":"Assembly of ascovirus HvAV-3h long DNA fragment using the Transformation-Associated Recombination (TAR) approach in yeast cells.","authors":"Heba A H Zaghloul, Zhengkun Xiao, Hengrui Hu, Guo-Hua Huang","doi":"10.1186/s12896-025-00964-8","DOIUrl":"10.1186/s12896-025-00964-8","url":null,"abstract":"<p><strong>Background: </strong>Synthetic biology is a young but rapidly growing field that allows for assembling long DNA fragments, including complete chromosomes. A key approach for long-DNA assembly is the Transformation Associated Recombination (TAR), which relies on efficient homologous recombination in yeast cells. Recent reports indicate that the TAR method efficiently assembles some human and animal viruses characterized by their large DNA genome size. The application of the TAR method to synthesize long DNA fragments derived from insect viruses is scarce. Therefore, this study aimed to explore the TAR approach for the construction of a long DNA fragment (>44.6 Kb) from the insecticidal Heliothesis virescens ascovirus 3h (HvAV-3h) dsDNA genome to assess the suitability of this approach in genome-wide engineering studies in this family of viruses.</p><p><strong>Results: </strong>The long DNA fragment assembly process involved three stages: first, we amplified 15 segments of about 2.9-3.2 Kb each via PCR. Next, we recombined these segments through three parallel TAR cycles, producing medium-sized fragments of about 15 Kb. Finally, we assembled these fragments in a single TAR cycle to form a long DNA fragment of about 44.6 kb. We identified some positive clones by colony PCR or restriction digestion pattern. To assess the quality of the assembled DNA fragment, we conducted next-generation sequencing (NGS). A comparative analysis of Sanger sequencing for medium-sized fragments and NGS data from the synthesized long-DNA fragment demonstrated a nearly matched mutation profile, suggesting that the identified mutations and deletions were present at initial synthesis. Both datasets aligned with the reference HvAV-3h strain, revealing three specific nucleotide mutations and three unique mutation regions.</p><p><strong>Conclusions: </strong>Overall, the in vivo TAR assembly method efficiently assembled a long DNA fragment derived from the ascovirus genome as a template. The process is cost-effective and can be scaled up to synthesize the entire genome for gene functional studies.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"42"},"PeriodicalIF":3.5,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12100921/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144126506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-22DOI: 10.1186/s12896-025-00979-1
Philomena Edet, Maurice Ekpenyong, Atim Asitok, David Ubi, Cecilia Echa, Uwamere Edeghor, Sylvester Antai
{"title":"Bio-induced overproduction of heterocycloanthracin-like bacteriocin in Lysinibacillus macroides by Aspergillus austroafricanus: optimization of medium conditions and evaluation of potential applications.","authors":"Philomena Edet, Maurice Ekpenyong, Atim Asitok, David Ubi, Cecilia Echa, Uwamere Edeghor, Sylvester Antai","doi":"10.1186/s12896-025-00979-1","DOIUrl":"10.1186/s12896-025-00979-1","url":null,"abstract":"<p><strong>Background: </strong>Plants and microorganisms are at the forefront of natural exploitable bioresources for the discovery of novel bioactive compounds (BACs) to provide solutions to food and agricultural challenges. The present study aimed to produce a novel biotechnologically-relevant BAC from a mangrove sediment bacterium under optimized bioprocess medium conditions. The BAC-producing bacteria were isolated via the crowded plate technique, and medium optimization was performed via sequential statistics of response surface methodology (RSM). The RSM model predictions were optimized, validated, and scaled up in a 5-L bioreactor via submerged batch fermentation. The BAC was extracted with ethyl acetate, purified via silica gel column chromatography, and identified via semipreparative high-performance liquid chromatography using bioactive standards with known retention times. The biocontrol, antioxidant and biopreservation potential of the BAC were evaluated via standard methods.</p><p><strong>Results: </strong>The results revealed that strain GKRMS-A9 produced the largest inhibition zone diameter (ZND) of 17 mm against the susceptible mould. The bacterium and its susceptible mould were identified as Lysinibacillus macroides and Aspergillus austroafricanus strains, respectively. Bioprocess medium optimization produced 9.6 g L<sup>- 1</sup> of the BAC with a ZND of 47.1 mm using 44.84% [v v<sup>- 1</sup>] rice processing effluent, 8.58 gL<sup>- 1</sup> casamino acid, 1.39 g L<sup>- 1</sup> MgSO<sub>4</sub>.7H<sub>2</sub>O, 2.78 g L<sup>- 1</sup> CaCl<sub>2</sub>.2H<sub>2</sub>O, 16.94% [v v<sup>- 1</sup>] inoculum volume, and 10.45 g L<sup>- 1</sup> Na<sub>2</sub>HPO<sub>4</sub>/NaH<sub>2</sub>PO<sub>4</sub>. The BAC concentration increased 48.7-fold in response to biological induction with susceptible mould. Silica gel chromatography revealed 9 bioactive fractions in the ethyl acetate extract, with fraction C (retention time of 9.02 min) eliciting the largest mean ZND of 38.1 ± 1.7 mm against Aspergillus austroafricanus. Fraction C was identified as a heterocycloanthracin-like class II bacteriocin with a molecular weight of 10.5 kDa.</p><p><strong>Conclusion: </strong>The bacteriocin 'macroidin' is stable over a wide range of pH values and temperatures and has significant antimicrobial activity against Gram-positive food-borne and phytopathogenic strains of bacteria and moulds. Its antioxidant activities against DPPH and ABTS*<sup>+</sup> radicals are comparable to those of ascorbic acid, making this biomolecule a promising agent for biopreservation and phytopathogen control applications in the food and agricultural sectors.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"41"},"PeriodicalIF":3.5,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12101020/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144126507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Fibromodulin-overexpressing fibroblast cells increase wound contraction, improve scar quality and enhance angiogenesis: an in-vivo study.","authors":"Negar Abdollahzadeh, Mehran Vatanchian, Fatemeh Oroojalian, Seyed Ehsan Enderami, Amir Amani, Reza Salarinia","doi":"10.1186/s12896-025-00975-5","DOIUrl":"10.1186/s12896-025-00975-5","url":null,"abstract":"<p><strong>Introduction: </strong>Fibromodulin, a small leucine rich proteoglycan has been suggested to have prominent role in wound healing. On the other hand, fibroblast cells, due to their ability to secrete growth factors and control inflammation in the wound area, have been proposed as effective approaches in cell therapy for wounds. In the current study we attempted to improve treatment results using a combination of fibroblast and fibromodulin features.</p><p><strong>Method: </strong>Fibroblast cells were isolated from the skin and transfected with a vector carrying the fibromodulin gene. Following the assessment of fibromodulin protein production, the effect of transfected fibroblast cells was studied in an animal wound model.</p><p><strong>Results: </strong>Flow cytometry analysis showed high expression of the CD90 marker (97.2%) and very low expression of the CD34 marker (0.47%). Additionally, enzyme-linked immunosorbent assay (ELISA) findings confirmed high expression of the fibromodulin gene in the transfected fibroblast cells. In vivo studies demonstrated that the animals treated with fibroblast cells transfected with fibromodulin (V + G+) exhibited significantly improved wound contraction on day 7 (i.e., contraction percentage: 21.79 ± 9.96%, compared with 7.23 ± 2.30% in the PBS-treated group). Histopathological studies also indicated improvements in angiogenesis score and collagen density score in the animals treated with the V + G + group.</p><p><strong>Conclusion: </strong>The results of this study showed that fibroblast cells expressing the fibromodulin gene improve wound contraction and some histological parameters in the deep wound model of the rat.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"40"},"PeriodicalIF":3.5,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12090437/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144101250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-17DOI: 10.1186/s12896-025-00977-3
Maysa M Ali, Somaya Nassar, Nivien Allam Nafady, Eman Mostafa Mohamed
{"title":"Bioremediation of malachite green dye toxicity under optimized conditions by Rhodotorula mucilaginosa AUMC13567.","authors":"Maysa M Ali, Somaya Nassar, Nivien Allam Nafady, Eman Mostafa Mohamed","doi":"10.1186/s12896-025-00977-3","DOIUrl":"10.1186/s12896-025-00977-3","url":null,"abstract":"<p><p>The escalating impacts of human urbanization and rapid industrial development have resulted in a growing global demand for synthetic textile dyes. Malachite green (MG) dye is a hazardous chromatic compound predominantly utilized as a textile dye, exhibiting a strong affinity for the substrate to which it is applied. When diluted, it serves as antimicrobial agent for fungal and bacterial diseases in fish farms. However, the disposal of textile dyeing wastes pose significant environmental challenges, contaminating soil, surface water, and adversely affecting human health and aquatic life. R. mucilaginosa exhibits robust growth and resistance capabilities even under conditions of nutrient deficiency or external stress. This study focuses on optimizing the malachite green dye degradation ability of the yeast strain Rhodotorula mucilaginosa AUMC13567. The results reveal that the optimal conditions for maximum decolorization which was 100% at 50 mg/L dye concentration involve cultivating R. mucilaginosa on medium comprising distilled water with 3% glucose, 0.5% peptone, 0.3% malt extract, and 0.3% yeast extract under aerobic conditions at 37˚C for 12 h at 150 rpm. Gas Chromatography-Mass Spectrometry (GC/MS) analysis confirms the yeast's efficacy in degrading toxic MG dye complex compounds into simple harmless metabolites. Fourier Transform Infrared (FTIR) analyses show formation of novel compounds post the degradation of MG dye. Further research is advised to identify and characterize the specific enzymes or metabolic pathways implicated in MG dye degradation by R. mucilaginosa AUMC13567. Cytotoxicity tests carried out on three human cell lines, including colorectal cancer, head and neck cancer, and healthy skin fibroblast, demonstrating that decolorized MG solution produces safe products devoid of cytotoxicity on tested human cell lines.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"39"},"PeriodicalIF":3.5,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12084967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144092653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-14DOI: 10.1186/s12896-025-00971-9
Ahmed H Elsaffany, Ahmed E M Abdelaziz, Abdullah A Zahra, Alsayed E Mekky
{"title":"Green synthesis of silver nanoparticles using cocoon extract of Bombyx mori L.: therapeutic potential in antibacterial, antioxidant, anti-inflammatory, and anti-tumor applications.","authors":"Ahmed H Elsaffany, Ahmed E M Abdelaziz, Abdullah A Zahra, Alsayed E Mekky","doi":"10.1186/s12896-025-00971-9","DOIUrl":"https://doi.org/10.1186/s12896-025-00971-9","url":null,"abstract":"<p><p>Bombyx mori silk is one of the most extensively studied types of silk due to its unique mechanical properties and biocompatibility, which have enabled its Utilization in medical applications Including surgical sutures since the second century. In the present study, a new method for the biosynthesis of silver nanoparticles (AgNPs) was explored by applying Bombyx mori cocoon extract as a sustainable and eco-friendly biological source. Unlike previous studies that primarily utilized plant or microbial extracts, this approach offers a more efficient alternative due to the unique protein and polyphenol content of silk cocoons, which enhances the stability and biological properties of the biosynthesized nanoparticles. The resulting AgNPs exhibited significant antibacterial, antioxidant, anti-inflammatory, and cytotoxic properties, opening new avenues for their therapeutic applications. This study expands the range of biological materials used in AgNP synthesis and provides deeper insight into how different bioactive components influence their functional properties. In this study, AgNPs were biosynthesized by mechanically processing extracted raw silk material with silver nitrate (AgNO₃). The synthesized nanoparticles were characterized by implementing several physicochemical techniques, including UV-visible spectrophotometry, FTIR, and XRD, and their morphology was examined through Transmission Electron Microscopy (TEM). The obtained AgNPs displayed a distinct absorption peak at 420 nm, with a particle size ranging between 5 and 25 nm, and displayed characteristic FTIR and XRD patterns typical of silver nanoparticles. The biosynthesized AgNPs demonstrated significant antimicrobial activity against Staphylococcus aureus (ATCC25923), Staphylococcus haemolyticus (ATCC29968), Escherichia coli (ATCC8739), and Klebsiella pneumoniae (ATCC2146). The antioxidant potential, assessed via the DPPH assay, yielded an IC50 value of 4.94 µg/ml, while the anti-inflammatory effect, evaluated using the membrane stabilization technique, showed an IC50 of 7.14 µg/ml. Additionally, AgNPs exhibited notable cytotoxic properties against Caco-2 and PANC1 cell lines, with IC50 values of 177.24 ± 2.01 µg/ml and 208.15 ± 2.79 µg/ml, respectively. Conversely, their impact on normal HFB-4 cells was minimal, with an IC50 of 582.33 ± 6.37 µg/ml, indicating a favorable safety profile. These observations highlight the multifunctional potential of silk-derived AgNPs, suggesting their applicability in various biomedical fields.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"38"},"PeriodicalIF":3.5,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12077009/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144075617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-14DOI: 10.1186/s12896-025-00974-6
Rozina Rashid, Uroosa Ejaz, Wissal Audah Alhilfi, Mohammed Alorabi, Syed Tariq Ali, Muhammad Sohail
{"title":"Bioconversion of wild Ipomoea pes-caprae and Suaeda fruticosa biomass: a novel application of thermostable xylanase from Neobacillus sedimentimangrovi.","authors":"Rozina Rashid, Uroosa Ejaz, Wissal Audah Alhilfi, Mohammed Alorabi, Syed Tariq Ali, Muhammad Sohail","doi":"10.1186/s12896-025-00974-6","DOIUrl":"https://doi.org/10.1186/s12896-025-00974-6","url":null,"abstract":"<p><p>Biomass from halophytes is considered as a promising chemical feedstock. Its bioconversion to obtain reducing sugars and to concomitantly improve antioxidant potential has been described less frequently. This is the first report describing application of xylanase from Neobacillus sedimentimangrovi for the saccharification of Ipomoea pes-caprae (IPC) and Suaeda fruticosa (SF). In this study, the biomass IPC and SF was separately or co-pretreated by freeze-thaw and 1% H<sub>2</sub>SO<sub>4</sub>. Results showed that significant amount of reducing sugar was obtained by saccharification of acid and freeze-thaw pretreated IPC (44 mg g<sup>- 1</sup>) and freeze-thaw pretreated SF (43 mg g<sup>- 1</sup>). The residues after saccharification were also analyzed for their antioxidant potential where IPC residues exhibited 1.13 folds higher potential than that of SF. Antioxidant potential (83.9%) was obtained when purified xylanase was used for the saccharification of IPC. Moreover, absence of lignin-related peaks in the NMR and IR spectra of the treated substrates indicated efficient delignification. The characteristic peaks of the hemicellulosic fractions in saccharified samples were also disturbed, indicating changes in the crystallinity of the substrates. The SEM images and spectra of the saccharified substrates clearly indicated the degradation of hemicellulosic content by xylanse.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"37"},"PeriodicalIF":3.5,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12076864/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144075613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-13DOI: 10.1186/s12896-025-00973-7
Fatma Gizem Avci, Tim Prasun, Volker F Wendisch
{"title":"Metabolic engineering for microbial production of sugar acids.","authors":"Fatma Gizem Avci, Tim Prasun, Volker F Wendisch","doi":"10.1186/s12896-025-00973-7","DOIUrl":"10.1186/s12896-025-00973-7","url":null,"abstract":"<p><p>Carbohydrates including sugar acids are commonly used as carbon sources in microbial biotechnology. These sugar acids are themselves desirable and often overlooked targets for biobased production since they find applications in a broad range of industries, examples include food, construction, medical, textile, and polymer industries. Different stages of oxidation for natural sugar acids can be distinguished. Oxidation of the aldehyde group yields aldonic acids, oxidation of the primary hydroxy group leads to uronic acids, and both oxidations combined yield aldaric acids. While the chemical oxidation of sugars to their acid forms often is a one-pot reaction under harsh conditions, their biosynthesis is much more delicate. Bio-based production can involve enzymatic conversion, whole-cell biotransformation, and fermentation. Generally, the in vivo approaches are preferred because they are less resource-intensive than enzymatic conversion. Metabolic engineering plays a crucial role in optimizing microbial strains for efficient sugar acid production. Strategies include pathway engineering to overexpress key enzymes involved in sugar oxidation, deletion of competing pathways to enhance the precursor availability and eliminate the product consumption, cofactor balancing for efficient redox reactions, and transporter engineering to facilitate precursor import or sugar acid export. Synthetic biology tools, such as CRISPR-Cas and dynamic regulatory circuits, have further improved strain development by enabling precise genetic modifications and adaptive control of metabolic fluxes. The usage of plant biomass hydrolysates for bio-based production further adds to the environmental friendliness of the in vivo approaches. This review highlights the different approaches for the production of C5 and C6 sugar acids, their applications, and their catabolism in microbes.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"36"},"PeriodicalIF":3.5,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12076931/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143952753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BMC BiotechnologyPub Date : 2025-05-12DOI: 10.1186/s12896-025-00970-w
Min Liu, Yanru Qiu, Erfu Xie, Pu Qian, Shuxian Yang, Simin Zhao, Wenjun Yan, Xuan Huang, Shuang Han
{"title":"Development of a chitosanase 3-like protein 1 assay kit and study of its application in patients with hepatocellular carcinoma.","authors":"Min Liu, Yanru Qiu, Erfu Xie, Pu Qian, Shuxian Yang, Simin Zhao, Wenjun Yan, Xuan Huang, Shuang Han","doi":"10.1186/s12896-025-00970-w","DOIUrl":"10.1186/s12896-025-00970-w","url":null,"abstract":"<p><strong>Objective: </strong>The detection kit for plasma Chitinase-3-like Protein 1 was developed using the magnetic bead chemiluminescence method, in order to investigate the diagnostic value of DD, FDP, CHI3L1, AFP-L3 and PIVKA-II in hepatocellular carcinoma.</p><p><strong>Method: </strong>The CHI3L1 detection kit was developed using the chemiluminescence method. The luminescence value obtained from the chemiluminescence analyzer was utilized for sensitive detection of CHI3L1, and the performance of the kit was evaluated accordingly. Moreover, this study enrolled 200 patients with hepatocellular carcinoma who were treated at the Oncology Department of the Affiliated Hospital of Jiangnan University between August 2022 and November 2023 as study subjects, while 100 healthy individuals undergoing physical examinations during the same period served as a control group. The plasma CHI3L1 levels in these subjects were measured using our institute's developed kit. Simultaneously, DD, FDP, AFP-L3, and PIVKA-II levels were assessed in all subjects to investigate their relationship with general pathology in patients with hepatocellular carcinoma. Additionally, ROC curves were generated to evaluate both single and combined detections' diagnostic efficacy for hepatocellular carcinoma.</p><p><strong>Result: </strong>The serological index changes of DD, FDP, AFP-L3, PIVKA-II, and CHI3L1 were not associated with patient gender. The concentrations of AFP-L3 and PIVKA-II in the 45-59 age group were significantly higher than in other groups (P < 0.05). Additionally, DD, CHI3L1, and PIVKA-II levels were markedly elevated in patients with tumors > 5 cm, medium-to-high differentiation, nerve invasion, lymph node metastasis, or distant metastasis. In advanced liver cancer (stages III-IV), DD, FDP, and CHI3L1 concentrations were significantly higher than in early-stage patients (stages I-II). For single diagnostic analysis, the AUC for CHI3L1 was 0.923, while the combined AUC for all five indices was 0.961, indicating greater diagnostic value when used together. The CHI3L1 chemiluminescence detection kit had a minimum detection limit of 1.50 ng/mL, with precision and accuracy within 10%, and R > 0.99. Compared to a clinical reference kit, the correlation coefficient (R) was 0.994, meeting clinical performance evaluation criteria.</p><p><strong>Conclusion: </strong>The CHI3L1 chemiluminescence kit developed meets clinical requirements. CHI3L1 can be used as an indicator for early screening of liver cancer, and the detection value of combined five indicators DD, FDP, AFP-L3, PIVKA-II and CHI3L1 is higher than that of single detection.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"35"},"PeriodicalIF":3.5,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12070687/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143964934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Hypoxia and HIF-1 inhibition enhance lentiviral transduction efficiency: a novel strategy for gene delivery optimization.","authors":"Qianyu Huo, Wentian Wang, Jiawen Dai, Xu Yuan, Dandan Yu, Bingqi Xu, Ying Chi, Huiyuan Li, Xiao Lei Pei, Guoqing Zhu, Lei Zhang","doi":"10.1186/s12896-025-00969-3","DOIUrl":"https://doi.org/10.1186/s12896-025-00969-3","url":null,"abstract":"<p><p>Lentiviral vectors are widely used for stable gene delivery, but their transduction efficiency can be limited by suboptimal experimental conditions. Here, we investigated the role of oxygen concentration and hypoxia-inducible factor 1 (HIF-1) signaling in lentiviral packaging and transduction. We found that packaging lentivirus under hypoxic conditions (10% O₂) significantly increased viral titers and transduction efficiency by approximately 10%. However, hypoxic conditions during viral entry impaired infection efficiency, likely due to HIF-1α-mediated cellular protective mechanisms. Pretreatment of cells with the HIF-1 inhibitor PX-478 reversed this effect, enhancing viral entry and genome integration in a dose-dependent manner. Combining hypoxic virus packaging with PX-478 pretreatment synergistically improved transduction efficiency by 20%. These findings suggest that HIF-1 inhibition and controlled hypoxia significantly enhance lentiviral transduction efficiency, establishing a versatile strategy with broad applicability across viral vector-dependent biomedical applications.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"34"},"PeriodicalIF":3.5,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12065270/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143966503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}