BMC Biotechnology最新文献

The study's goals are to fabricate PLGA nanoparticles (PNPs) loaded with apigenin (AP) and adenosine (AD) using a microfluidic preparation method to a standard emulsification method and investigate the possible heart-protective effects of AP-AD PNPs made using the emulsification method. Compared to microfluidics, the emulsification method fabricated small-size nanoparticles, which are better at encapsulating drugs, retaining more drugs, and having a low viscosity for the myocardial infarction (MI) injection. TheMI model was developed using SD rats injected under the skin with 85 mg/kg doxorubicin (DOX) for 2 days. The metabolic results showed that our AP-AD PNPs accelerated the blood flow in rats with MI, which increased the amounts of AP and AD in the circulatory system. This led to significant improvements in the cardiac index and lower amounts of AST, LDH, and CK in the blood. A histopathological study using Hematoxylin&eosin, and TUNEL staining showed that cardiac function had improved and apoptosis had decreased. Moreover, tests that checked the amounts of IL-6, TNF-α, NO, GSH, MDA, and SOD showed that AP-AD PNPs may help treat MI by reducing oxidative stress and inflammation, making it a potentially useful therapeutic approach.

This work examined the antibacterial and antibiofilm properties of alginate/collagen nanoparticles containing limonene. The multi-drug resistant (MDR) strains were screened, and the morphological features of the produced nanoparticles were determined utilizing SEM, DLS, and FTIR. Additionally, the encapsulation effectiveness, stability, and drug release were assessed. The levels of OmpA and Bap biofilm genes were assessed using qRT-PCR. At the same time, the antibacterial and cytotoxic activities of the nanoparticles were evaluated using well diffusion and MTT techniques, respectively. LAC nanoparticles measuring 300 ± 9.6 nm in size, 83.64 ± 0.19% encapsulation efficiency, and 60-day stability at 4 °C were synthesized. The biological investigation demonstrated that LAC nanoparticles had potent antibacterial capabilities. This was shown by their ability to significantly decrease the transcription of OmpA and Bap biofilm genes at a statistically significant level of p ≤ 0.05. The nanoparticles exhibited reduced antibiotic resistance compared to free limonene and alginate/collagen. Compared to limonene, LAC nanoparticles exhibited negligible cytotoxicity against HEK-293 at doses ranging from 1.56 to 100 µg/mL (p ≤ 0.01). The findings underscore the potential of LAC nanoparticles as a breakthrough in the fight against highly resistant pathogens. The potent antibacterial effects of LAC nanoparticles versus Acinetobacter baumannii (A. baumannii) MDR strains, considered highly resistant pathogens of significant concern, could inspire new strategies in antibacterial research.

Background: Natural pigments are becoming more significant because of the rising cost of raw materials, pollution, and the complexity of synthetic pigments. Compared to synthetic pigments, natural pigments exhibit antimicrobial properties and is less allergic. Pigments from microbial sources could easily be obtained in an inexpensive culture media, produced in high yields, and microbes are capable of producing different colored pigments. Searching for new sources for natural pigments to replace synthetic ones in food applications has become an urgent necessity, but the instability of these compounds is sometimes considered one of the obstacles that reduce their application. Encapsulation provides an ideal solution for natural dye protection through a controlled release strategy. Thus, this study aims at isolation of several soil fungi and subsequent screening their pigment production ability. The chosen pigment-producing fungal strain underwent full identification. The produced pigment was extracted with ethyl acetate and estimated spectrophotometrically. As there is a necessity to obtain a high pigment yield for efficient industrial application, the best production medium was tested, optimum conditions for maximum dye production were also investigated through the response surface methodology, and gamma irradiation was also employed to enhance the fungal productivity. Encapsulation of the produced pigment into chitosan microsphere was tested. The pigment release under different pH conditions was also investigated.

Results: A new strain, Fusarium verticillioides AUMC 15934 was chosen and identified for a violet pigment production process. Out of four different media studied, the tested strain grew well on potato dextrose broth medium. Optimum conditions are initial medium pH 8, 25 °C-incubation temperature, and for 15-day incubation period under shaking state. Moreover, a 400 Gy irradiation dose enhanced the pigment production. Chitosan microsphere loaded by the pigment was successfully prepared and characterized by infrared spectroscopy and scanning electron microscopy.

Conclusion: This irradiated Fusarium strain provides a more economically favorable source for production of a natural violet dye with an optimum productivity, enhanced yield, and improved properties (such as, enhanced stability, controlled release, and bioaccessibility) by encapsulation with chitosan for efficient application in food industry.

Synthetic dyes, such as Alizarin Red S, contribute significantly to environmental pollution. This study investigates the biosorption potential of Alhagi maurorum biosorbent for the removal of Alizarin Red S (ARS) from aqueous solutions. Fourier transform infrared spectroscopy (FTIR) was used to analyze the biosorbent's adsorption sites. Various parameters were optimized to maximize dye adsorption. An optimal removal efficiency of 82.26% was attained by employing 0.9 g of biosorbent with a 25 ppm dye concentration at pH 6 and 60 °C over 30 min. The data were modeled using various isothermal and kinetic models to understand the adsorption behavior. Thermodynamic parameters indicated that the adsorption process was spontaneous and endothermic. The pseudo-second-order kinetic model best described the data, indicating chemisorption as the rate-limiting step. The data matched best to the Langmuir model, indicating that the adsorption occurs as a monolayer on uniform surfaces with a finite number of binding sites. The model showed a strong correlation (R² = 0.991) and a maximum adsorption capacity (q_max) of 8.203 mg/g. Principal component analysis (PCA) identified temperature as the dominant factor, with the primary component, PC1 capturing 100% of its effect. The mechanisms involved in ARS biosorption on A. maurorum include electrostatic interactions, hydrogen bonding, hydrophobic interactions, dipole-dipole interactions, and π-π stacking. Alhagi maurorum showed promising potential for biosorbing toxic dyes from contaminated water, suggesting further investigation for practical applications.

Optimizing extraction conditions can help maximize the efficiency and yield of the extraction process while minimizing negative impacts on the environment and human health. For the purpose of the current study, an artificial neural network (ANN) combined with a genetic algorithm (GA) was utilized for that the extraction conditions of Hypericum spectabile were optimized. In this particular investigation, the main objective was to get the highest possible levels of total antioxidant status (TAS) for the extracts that were obtained. In addition to this, conditions of the extract that exhibited the maximum activity have been determined and the biological activity of the extract that was obtained under these conditions was analyzed. TAS values were obtained from extracts obtained using extraction temperatures of 30-60 °C, extraction times of 4-10 h, and extract concentrations of 0.25-2 mg/mL. The best model selected from the established ANN models had a mean absolute percentage error (MAPE) value of 0.643%, a mean squared error (MSE) value of 0.004, and a correlation coefficient (R) value of 0.996, respectively. The genetic algorithm proposed optimal extraction conditions of an extraction temperature of 59.391 °C, an extraction time of 8.841 h, and an extraction concentration of 1.951 mg/mL. It was concluded that the integration of ANN-GA can successfully be used to optimize extraction parameters of Hypericum spectabile. The total antioxidant value of the extract obtained under optimum conditions was determined as 9.306 ± 0.080 mmol/L, total oxidant value as 13.065 ± 0.112 µmol/L, oxidative stress index as 0.140 ± 0.001. Total phenolic content (TPC) was 109.34 ± 1.29 mg/g, total flavonoid content (TFC) was measured as 148.34 ± 1.48 mg/g. Anti-AChE value was determined as 30.68 ± 0.77 µg/mL, anti-BChE value was determined as 41.30 ± 0.48 µg/mL. It was also observed that the extract exhibited strong antiproliferative activities depending on the increase in concentration. As a result of LC-MS/MS analysis of the extract produced under optimum conditions in terms of phenolic content. The presence of fumaric, gallic, protocatechuic, 4-hydroxybenzoic, caffeic, 2-hydoxycinamic acids, quercetin and kaempferol was detected. As a result, it was determined that the H. spectabile extract produced under optimum conditions had significant effects in terms of biological activity.

Cloning is a key molecular biology procedure for obtaining a genetically homogenous population of organisms or cell lines. It requires the expansion of new cell populations starting from single genetically modified cells. Despite the technical progress, cloning of many cell lines remains difficult. Cloning often fails either due to the strenuous conditions associated with manipulating cells or because many cells don't tolerate a single-cell state. Here we describe a new cloning method utilizing low adhesion microcavity plates. This new technique, named microcavity-assisted cloning (MAC) was developed to clone difficult-to-clone HepG2 cells. The clones were produced following CRISPR/Cas9 knockout of the GSTA1 gene by a random distribution of 200, 400, and 800 cells into 550 microcavities of a 24-well low adhesion plate originally designed for the culture of spheroids. The knockout of GSTA1 was verified at the protein level using Western blotting. The advantages of the MAC method are its low cost, ease of the procedure, and the possibility of scaling up the throughput and automatization.

Background: Hepatitis B virus (HBV) clearance depends on an effective adaptive immune response, especially HBV-specific T cell-mediated cellular immunity; however, it is difficult to produce enough HBV-specific T cells effectively.

Results: In this work, we investigated the proportions of stimulated cells, serum, and culture media as the three primary factors to determine the most effective procedure and applied it to HLA-A2 (+) people. In parallel, we also examined the correlation between clinical parameters and HBV-specific immunity. Concerning amplification efficiency, 4 × 10⁵ cells stimulation was superior to 2 × 10⁶ cells stimulation, AIM-V medium outperformed 1640 medium, and fetal bovine serum (FBS) exceeded human AB serum under comparable conditions. As expected, this procedure is also suitable for developing HBV-specific CD8 + T cells in HLA-A2(+) individuals. Expanded HBV-specific T cell responses decreased with treatment time and were negatively correlated with HBV DNA and HBsAg. Furthermore, the number of HBV-specific IFN-γ + SFCs was strongly correlated with the ALT level and negatively correlated with the absolute lymphocyte count and the ALB concentration.

Conclusions: We confirm that stimulating 4 × 10⁵ PBMCs in AIM-V medium supplemented with 10% FBS is the best approach and that HBeAg, HBsAg, and ALB are independent predictors of HBV-specific T-cell responses.

Sepsis is an inevitable stage of bacterial invasion characterized by an uncontrolled inflammatory response resulting in a syndrome of multiorgan dysfunction. Most conventional antibiotics used to treat sepsis are efficacious, but they have undesirable side effects. The green synthesised Ag NPs were synthesized by 5 g of the earthworm extract dissolved in a volume of 500mL of distilled water and then added to 2,500 mL aqueous solution of 1mM silver nitrate at 40 °C. After 4 h, the mixture was then allowed to dry overnight at 60 °C. Later, Ag NPs were washed and collected. They were characterized by X-ray diffraction, ultraviolet-visible spectroscopy, and transmission electron microscopy. Sepsis model as induced by feces-intraperitoneal injection method. Eighteen male mice were assigned into three main groups: the control group, the sepsis-model group, and the Ag NPs-treated group. The control group received a single oral dose of distilled water and, after two days, intraperitoneally injected with 30% glycerol in phosphate buffer saline. The Sepsis-model group received a single oral dose of distilled water. Ag NPs - The treated group received a single oral dose of 5.5 mg/kg of Ag NPs. After two days, the sepsis-model group and Ag NPs-treated group were intraperitoneally injected with 200 µL of faecal slurry. Ag NPs treatment in septic mice significantly decreased liver enzyme activities, total protein, and serum albumin. Moreover, Ag NPs significantly enhanced kidney function, as indicated by a significant decrease in the levels of creatinine, urea, and uric acid. In addition, Ag NPs showed a powerful antioxidant effect via the considerable reduction of malondialdehyde and nitric oxide levels and the increase in antioxidant content. The histopathological investigation showed clear improvement in hepatic and kidney architecture. Our findings demonstrate the protective efficacy of biogenic Ag NPs against sepsis-induced liver and kidney damage.

Wound infections resulting from pathogen infiltration pose a significant challenge in healthcare settings and everyday life. When the skin barrier is compromised due to injuries, surgeries, or chronic conditions, pathogens such as bacteria, fungi, and viruses can enter the body, leading to infections. These infections can range from mild to severe, causing discomfort, delayed healing, and, in some cases, life-threatening complications. Zinc oxide (ZnO) nanoparticles (NPs) have been widely recognized for their antimicrobial and wound healing properties, while cinnamic acid is known for its antioxidant and anti-inflammatory activities. Based on these properties, the combination of ZnO NPs with cinnamic acid (CA) was hypothesized to have enhanced efficacy in addressing wound infections and promoting healing. This study aimed to synthesize and evaluate the potential of ZnO-CN NPs as a multifunctional agent for wound treatment. ZnO-CN NPs were synthesized and characterized using key techniques to confirm their structure and composition. The antioxidant and anti-inflammatory potential of ZnO-CN NPs was evaluated through standard in vitro assays, demonstrating strong free radical scavenging and inhibition of protein denaturation. The antimicrobial activity of the nanoparticles was tested against common wound pathogens, revealing effective inhibition at a minimal concentration. A zebrafish wound healing model was employed to assess both the safety and therapeutic efficacy of the nanoparticles, showing no toxicity at tested concentrations and facilitating faster wound closure. Additionally, pro-inflammatory cytokine gene expression was analyzed to understand the role of ZnO-CN NPs in wound healing mechanisms. In conclusion, ZnO-CN NPs demonstrate potent antioxidant, anti-inflammatory, and antimicrobial properties, making them promising candidates for wound treatment. Given their multifunctional properties and non-toxicity at tested concentrations, ZnO-CN NPs hold significant potential as a therapeutic agent for clinical wound management, warranting further investigation in human models.