Electronic Journal of Biotechnology

circUBAP2 ameliorates hypoxia-induced acute myocardial injury by competing with miR-148b-3p and mediating CDKN1B expression

FeiFei Li — 2024-03-15

Background: A recent high-throughput sequencing study revealed an anomalous underexpression of circular RNA UBAP2 (circUBAP2) in acute myocardial infarction (AMI), yet its biological function within this context remains elusive. This study aims to unravel whether circUBAP2 is instrumental in modulating the pathogenesis of AMI and to illuminate the underlying molecular mechanisms at play.

Results: circUBAP2 was abnormally low expressed in AMI. Inducing circUBAP2 ameliorated hypoxia-induced myocardial cell injury by enhancing cellular viability, and decreasing lactate dehydrogenase release, apoptosis, inflammation, and oxidative damage. circUBAP2 targeted miR-148b-3p, miR-148b-3p overexpression offset circUBAP2-induced cardioprotection. Cyclin-dependent kinase inhibitor 1B (CDKN1B) was mediated by miR-148b-3p, and CDKN1B upregulation suppressed the deleterious effect of circUBAP2 silencing on hypoxic AC16 cells. In addition, overexpression of circUBAP2 improved myocardial injury, decreased myocardial cell apoptosis, and alleviated inflammation and oxidative stress in AMI mice.

Conclusions: circUBAP2 ameliorates AMI by competitively binding to miR-148b-3p and mediating CDKN1B expression.

Synthesis of hot spring origin bacterial cell wall polysaccharide-based copper nanoparticles with antibacterial property

Aparna Banerjee — 2024-03-15

Background: At present, research on facile, green synthesis of nanoparticles has significantly increased because of its fast, one-step, cost-effective, time-efficient, and non-toxic nature. In this study, we have reported a single-step green synthesis of copper nanoparticles using cell wall polysaccharides of a hot spring origin, thermotolerant Bacillus species.

Result: Copper nanoparticles were characterized using UV-visible spectrophotometry, fluorescence and Fourier transform infrared spectroscopy, scanning electron microscopy with energy dispersive spectroscopy, particle size, and zeta potential analyses. UV-visible spectra of synthesized copper nanoparticles exhibited a band cantered between 220–235 nm, characteristic spectra of copper oxide nanoparticles. Infrared spectra showed the band at 490-530 cm⁻¹ corresponding to metal-oxygen or copper nanoparticle vibration, supporting the presence of copper oxide nanoparticles in the monoclinic phase. The energy dispersive spectra of copper nanoparticles exhibited a strong signal from elemental copper. The dynamic Light Scattering pattern confirmed the nanoparticle nature of the studied sample. These nanoparticles showed preferential activity against gram-negative pathogens, Salmonella typhi and Escherichia coli. The thermodynamic nature of the nanoparticles is also established for its antibacterial actions.

Conclusions: The antibacterial action and its thermodynamics reinforce the possible use of copper nanoparticles as an alternative to commercially available antimicrobials. This study may open a new path for future studies to treat harmful microorganisms resistant to traditional antibiotics in a greener way.

A novel nanoemulsion based on clove and thyme essential oils: Characterization, antibacterial, antibiofilm and anticancer activities

Amr H. Hashem — 2024-03-15

Background: Essential oil nanoemulsions have received much attention in the last period due to their ability to fight microbes and cancers. In the current study, clove and thyme essential oils CL+TH-emulsion and CL+TH-nanoemulsion were prepared through an eco-friendly method. The prepared CL+TH-nanoemulsion was characterized using DLS and TEM analyses.

Results: Results revealed that CL+TH-nanoemulsion droplets were spherical in shape and nanoform in size (68.6 nm) with PDI 0.281. MIC concentrations of CL+TH-nanoemulsion against tested bacteria were found to be between 6.25 and 25 mg/mL. After being exposed to MICs of CL+TH-emulsion and CL+TH-nanoemulsion, which additionally prompted 1.43 log and 3.12 log declines, accordingly, as opposed to untreated (Control), the number of cells grown in the generated biofilms decreased. Furthermore, CL+TH-nanoemulsion exhibited anticancer activity more than CL+TH-emulsion toward HepG2 and MCF-7. Also, the effect of CL+TH-nanoemulsion is more effective and significantly cytotoxic than taxol on MCF-7. Besides, both prepared emulsions increased the rate of apoptosis and decreased the cell viability % of MCF-7 by increasing the activity of caspases 8 and 9. Moreover, CL+TH-nano emulsion decreased the activity of VEGFR-2 in MCF-7 in a more pronounced manner than CL+TH-emulsion and taxol.

Conclusions: The prepared CL+TH-nanoemulsion had antibacterial, and antibiofilm as well as anticancer properties, which can be used in different biomedical applications after extensive studies in vivo.

Circular RNA FMN2 motivates colorectal cancer development by mediating tumor-associated macrophage polarization by controlling the microRNA-150-5p/PIK3R3 axis

YongYi Cao — 2024-03-15

Background: It has been reported that circular RNA formin 2 (circFMN2) can motivate colorectal cancer (CRC) proliferation. However, the effect of circFMN2 on the polarization of tumor-associated macrophages (TAMs) in the CRC tumor microenvironment remains unclear. The study was to figure out the latent mechanism by which circFMN2 impacts TAM polarization to motivate the malignant behavior of CRC cells.

Results: circFMN2 and PIK3R3 levels were reduced in M1 macrophages but elevated in M2 macrophages, whereas miR-150-5p level was the opposite. circFMN2 knockdown downregulated M2 macrophage markers CD163, CCL22 and CD206 and upregulated M1 macrophage markers CD86, TNF-α and IL-1β in M2 macrophages. Co-culture with M2 macrophage-conditioned medium with circFMN2 knockdown reduced CRC proliferation, invasion, and migration, while knockdown of miR-150-5p had the opposite effect. CircFMN2 adsorbed miR-150-5p to mediate PIK3R3 in M2 macrophages. Overexpression of miR-150-5p can reverse the promoting effects of overexpression of circFMN2 on M2 polarization, CRC cell proliferation, invasion, and migration. Elevation of PIK3R3 could turn around the repressive effect of circFMN2 knockdown on M2 polarization and CRC cell proliferation, invasion, and migration. In an in vivo model, M2 macrophages expressing low or high circFMN2 were co-transplanted with CRC cells into nude mice, resulting in inhibition and promotion of tumor growth, respectively.

Conclusions: All in all, circFMN2 mediates TAM polarization to M2 type by controlling the miR-150-5p/PIK3R3 axis to motivate CRC development and may offer a latent molecular target for CRC treatment.

Successful CRISPR/Cas9-mediated HDR at individual DNA breakpoints using TFO-based targeted template design

Zahra Ebrahimi — 2024-03-15

Background: Targeted insertion of the repair template into the genome is a common strategy for high-precision base replacements; however, the main challenge likely remains regarding the limited efficiency of homologous-directed repair (HDR). A precise genome cut achieved by CRISPR-Cas9 system combining with a single-stranded oligodeoxynucleotide (ssODN), as the donor template, improves significantly the rate of HDR. It is well-established that the spatial availability of the donor template to the repair system effectively enhances knock-in events in CRISPR-Cas9. PolyPurine Reverse Hoogsteen hairpins (PPRHs), as an alternative repairing strategy, benefits from a Triplex-forming oligonucleotide (TFO) for the repair template providing the ease of access. The main objective of the study was to evaluate the HDR frequency as a result of improvement of the spatial accessibility of the donor template adjacent to the cutting site. Hence, a flanking purine-rich hairpin complementary to the genomic DNA adjacent to the repairing site was fused to the ssODN with the incorporated bases for the alteration of EGFP to EBFP.

Results: Results from the comparison between the donor templates, ssODN and TFO-tailed ssODN, demonstrated an increased rate of knock-in from 18.2% ± 1.09 to 38.3% ± 4.54, respectively. From another perspective, findings indicated that the targeted Cas9-mediated DNA cleavage improves the efficiency of the repair-PPRH approach four-fold, as well.

Conclusions: The present study provides a viewpoint that highlights the significance of the designing of the donor template in terms of the structural features and positional access for the HDR-based repairing CRISPR-Cas9 systems.

Enzymatically acylated naringin with gut modulation potential

Elisa Gutiérrez-Navarro — 2024-03-15

Background: Naringin is one of the main flavonoids in citrus fruits and byproducts. This flavanone has been shown to be a good antioxidant nutraceutical component, and it also has potential as a gut microbiome modulator, although its applications in final formulations represent a challenge due to its low solubility, both in water and in organic solvents. This work addresses this problem by functionalizing naringin through enzymatic acylation.

Results: The enzymatic acylation catalyzed by the lipase Novozym® 435 and using acyl donors of different chain lengths, acetate (C2), propionate (C3), and laurate (C12), yielded in conversions of 95% at 24 h and 100% at 48 h, generating a monoacylated product. Both the aqueous and solvent solubility of acylated naringin products were improved while maintaining or even increasing their antioxidant activity.

Conclusions: This acylation process significantly enhanced both the water and solvent solubility of the acylated naringin products while preserving or even enhancing their antioxidant activity. In addition to the gut-modulating properties of flavonoids, acylating them with short- and medium-chain fatty acids could enhance their potential applications in the emerging field of research dedicated to understanding and modulating gut health.

Screening for Aspergillus fumigatus strain-2T-2 with high chitosanase production activity and its application in chitosan degradation

Haoyue Yang — 2023-03-15

Background: High chitosanase-producing microorganisms from natural sources have extensive applications in food and agriculture. This study aimed to optimal conditions for high-activity chitosanase production. A named CGMCC21422 chitosanase-producing strain -2T-2 was isolated from soil and identified named as Aspergillus fumigatus chitosanase (A. fumigatus chitosanase). This enzymatic activity was validated in various culture conditions. It is stored in the China General Microbiological Culture Collection Center. The efficacy of A. fumigatus chitosanase in the degradation of chitosan was validated.

Results: In this study, we determined that the optimal fermentation conditions of stain-2T-2 were 1.0% powered chitosan, 0.8% ammonium nitrate, 37°C culture temperature, initial pH 5.0, culture time 6 d, bottle volume 50 mL, and 2% inoculation dosage. Under these culture conditions, the highest enzyme activity of fermentation broth in the shaker flask reached 827.53 U/mL. The optimal reactive conditions of A. fumigatus-produced chitosanase are 55–60°C and pH 4.5. When the reactive temperature was over 60°C, the A. fumigatus chitosanase was easily inactivated. The chitosanase catalyzed substrate chitosan to produced ≈20% chito-oligosaccharide and ≥80% glucosamine salt samples in a variety of acidic solutions. These reactive products are not cytotoxic or mild to MH7A cells.

Conclusions: A. fumigatus chitosanase strain -2T-2 is a strain with high chitosanase and can catalyze chitosan into chito-oligosaccharide in acidic solutions.

In silico and in vitro assessment of the anti-β-amyloid aggregation and anti-cholinesterase activities of Ptaeroxylon obliquum and Bauhinia bowkeri extracts

Michael C. Ojo — 2023-03-15

Background: Alzheimer’s disease (AD) is the most common form of dementia and dementia constitutes the fifth leading cause of mortality across the globe. Available treatment modalities and drugs have abysmally failed to curtail AD. This study evaluated the mitigation of Aβ aggregation and anti-cholinesterase activities with the crude extracts of Ptaeroxylon obliquum and Bauhinia bowkeri. Computational studies of the most abundant phytochemicals from the crude extracts of both plants with proteins were investigated. The phytochemical composition of the different crude extracts (hexane, DCM, and ethanol) of the plants were analyzed with FTIR and GC-MS. The inhibitory potential of the extracts on BACE-1 and cholinesterase activities was determined with both computational molecular docking studies and in vitro enzyme assays. Their anti-aggregation properties were confirmed with Thioflavin-T assay and TEM.

Results: The in silico studies revealed that though thunbergol and cyclotetradecatriene (the major constituents of the extracts) inhibited all the proteins, the latter exhibited the best inhibitory potential. The in vitro results showed that while the dichloromethane (DCM) extract of P. obliquum had the highest butyrylcholinesterase (BuChE) inhibitory activity (1.77 µg/ml), the hexane and ethanol extract of B. bowkeri exhibited the highest β-site amyloid precursor cleaving enzymes-1 (BACE-1) (30.4 µg/ml) and acetylcholinesterase (AChE) (58.11 µg/ml) inhibitory efficacy, respectively. The ethanol extract (160 μg/ml) of B. bowkeri had the most efficacious anti-aggregation activity.

Conclusions: This study suggests that the plants could possess neuroprotective effects and could also be sources of anti-AD novel drugs.

Low-alcohol light beer enriched with olive leaves extract: Cold mashing technique associated with interrupted fermentation in the brewing process

Eliziane Cappelin — 2023-03-15

Background: Beer is the most consumed alcoholic beverage globally, and the demand for differentiated beers with peculiar characteristics has intensified among beer consumers, creating a significant market niche. In this study, we developed a low alcohol light craft beer enriched with olive leaf extract (Olea europaea L.). The cold mashing technique associated with interrupted fermentation was used in the mashing step. Different concentrations of olive leaf extract (0.5, 1.0 and 2.0%) were added at the maturation stage. The samples were characterized by physicochemical parameters, phenolic and polyphenolic content, bioactive compounds, antioxidant potential, and microbiological quality.

Results: The cold mash technique associated with interrupted fermentation provided a low-alcohol beer (≅1.3%). The bitterness dimension (19.0 to 23.2 IBU) and color (9–17 EBC) parameter were in accordance with the Beer Judge Certification Program (BJCP) for the American Blond Ale-style. The addition of the extract enriched the content of total phenolics (171.09 to 437.4 mg GAE/mL) and polyphenolic (221.4 to 729.0 mg/L). Coumaric, ferulic, and cinnamic phenolic acids were detected in appreciable amounts in the beers. Oleuropein was the major compound in the beverage and plant extract. After adding 2% extract, the ABTS and DPPH radical scavenging activity, as well as the ferric reduction power, increased in beers by 28.4%, 449.1%, and 120.5%, respectively.

Conclusions: The extract of O. europaea L. promoted the enrichment of low-alcohol beer samples with bioactive compounds and antioxidant potential. The results obtained indicated the potential use of O. europaea L. extract as a natural oxidant in other beverages and food products.