Recent Patents on Biotechnology - Online First

Urinary Tract Infections (UTIs) caused by Escherichia coli are a major concern due to rising antimicrobial resistance. Long non-coding RNAs (lncRNAs) play crucial roles in gene regulation, but their involvement in UTIs and their implication in antimicrobial resistance mechanisms are not well understood. This study investigates the association between specific long non-coding RNAs, immune response, and antibiotic resistance in patients with infections caused by E. coli.

Quantification of specific lncRNAs (NEAT1-1, NEAT1-2, MIR3142HG, AK170409, and IL7AS) was performed using quantitative PCR. Inflammatory markers IL-1β, IRF3, and NF-κB were measured in UTI patients using ELISA kits to assess their biological response. The minimum inhibitory concentration of eleven antibiotics was determined by testing all 25 urine samples and categorized as sensitive, intermediate, or resistant.

NEAT1-1, NEAT1-2, MIR3142HG, and AK170409 were significantly upregulated (p < 0.05). Inflammatory markers were significantly elevated in all samples: IL-1β at 72.36±13.8pg/mL, IRF3 at 79.36±15.01pg/mL, and NF-κB at 4.43±0.82pg/mL

(p < 0.0001). NEAT1-2 and AK170409 expression correlated with distinct antibiotic response patterns (p < 0.05). Differences in biological and hematological data were observed among UTI patients with varying topographic expressions of specific long non-coding RNAs (lncRNAs).

Specific lncRNAs may be involved in modulating immune responses and influencing antibiotic susceptibility in UTIs. Their expression patterns reflect both the severity of infection and resistance profiles, suggesting a functional role in the development of antimicrobial resistance.

LncRNAs are potential biomarkers for UTIs caused by E. coli. Future studies should focus on elucidating their role in the development of antibiotic resistance and exploring their patentable applications in biotechnology.

Polycyclic aromatic hydrocarbons (PAHs) are toxic petroleum byproducts in soil, exhibiting significant genotoxic properties. Microorganisms residing in contaminated soils serve as effective detoxifying agents. Among various strategies, bioremediation is an efficient biological method for detoxifying PAHs.

Hundreds of soil samples were collected from the Gulf of Suez, Egypt. The isolation process utilized an enrichment culture system with phenol naphthalene (PN) (10 mg/mL) as the primary carbon source. HPLC analysis was applied to confirm PN degradation. Consequently, the bacterial strain was characterized morphologically, biochemically, and through partial sequencing of its 16S rRNA gene. Subsequently, its plasmid was purified to transfer its phenotype to Escherichia coli. Finally, a bioremediation approach was conducted to test its PAH degradation.

HPLC analysis was performed to confirm PN degradation by the isolated strain. The isolated strain was identified as Lysinibacillus species AAS1 (OR044755.1) with 98.43% sequence similarity to the Lysinibacillus genus. Subsequently, E. coli transformants with the isolated plasmid were grown in the presence of PN as the primary carbon source. Finally, the bioremediation assay of the isolated strain exhibited a high efficiency in detoxifying PN.

The novel identified Lysinibacillus species AAS1 (OR044755.1) shows promise for PAHs detoxification, which may lead to the exploration of a biological agent for the remediation of water, soil contaminated with PAHs and patents.

A novel bacterial strain bearing a plasmid that can degrade PN was isolated from Egyptian petroleum waste-contaminated soil. It paved the way for further studies to isolate the whole gene(s) responsible for such degradation.

Biofilm production is a key factor in the development of antibiotic resistance in multidrug-resistant Klebsiella pneumoniae (K. pneumoniae), a significant contributor to healthcare-associated infections (HAIs). Kaempferol, a flavonoid, is widely recognized for its ability to combat various microorganisms.

Our goal is to assess the impact of kaempferol on K. pneumoniae biofilms by determining the level of gene expression for the biofilm-forming genes.

Fifty K. pneumoniae isolates were studied. Different doses of kaempferol with a concentration range of 0.04 to 100% in Luria Bertani broth (LB) medium were incubated at 37°C for 24 h with forty-three K. pneumoniae strong and intermediate biofilm producers. The minimum inhibitory concentration (MIC) of kaempferol was determined. Molecular detection of the biofilm-forming genes (mrkA, pgaA, wbbM, and wzm) was performed on all isolates before and after kaempferol treatment at 0.5 x MIC.

Seven isolates out of 50 (14%) exhibited weak biofilm formation ability, 6 out of 50 (12%) were moderate producers, and 37 out of 50 (74%) were strong producers. The MIC values of kaempferol for K. pneumoniae ranged from 50% to 6.25% (p = 0.0003). The levels of expression of the studied genes were slightly decreased after treatment compared with their corresponding values before treatment.

Kaempferol has shown potential in disrupting biofilms by inhibiting key genes (mrkA, pgaA, wbbM, wzm) involved in adhesion and biofilm matrix synthesis, although its effect is moderate. In vitro testing revealed that kaempferol inhibits biofilm formation at varying concentrations depending on the bacterial strain, with gene expression downregulation indicating its interference in biofilm-related pathways. Patent related to this topic has been mentioned along the text.

Based on current knowledge, few research studies have investigated the impact of kaempferol on K. pneumoniae biofilms. Our results show that its effect on the biofilms of this bacterium is moderate to weak. Further research is necessary to determine potential synergies with other treatments.

Medicines and herbal formulations are derived from different parts of medicinal plants, which are the best-known sources for treating various diseases. This research focuses on assessing the antimicrobial potential of crude extracts from the leaves and roots of Cenchrus biflorus Roxb.

Methanol, hydroethanol (50:50), and aqueous extracts were obtained using the Soxhlet extraction method. The disc diffusion method was used to study the antimicrobial activity of the extracts against a variety of test microorganisms, including bacteria (Escherichia coli and Bacillus subtilis) and fungus (Aspergillus niger). The disc diffusion method was used to assess bacterial susceptibility, revealing the potent inhibitory effect of the methanol extract on E. coli. All extracts demonstrated significant antimicrobial activity against various microorganisms.

Remarkably, methanol extract of leaf demonstrated the highest antibacterial activity, with a 16.3 ± 1.78 mm zone of inhibition (ZOI) with Activity Index (AI) of 0.875, and a Relative Percentage Inhibition (RPI) of 80 against E. coli, followed by Bacillus subtilis (ZOI = 15.5 ± 1.31 mm, AI = 0.869, RPI = 78.57). The methanol extract of the root showed strong antifungal activity against Aspergillus niger (with a 12.9 ±1 mm ZOI, AI = 0.636, and RPI = 42.85), while the water extract of the root displayed 7.8 mm inhibition zones.

Methanol and hydroethanol extracts of the leaf and root exhibited strong inhibitory effects against selected microbial strains. Each plant solvent extract suppressed microbial development in a distinct manner, and methanol and hydroethanol extracts inhibited microbial development more efficiently than aqueous extracts. Interestingly, water extracts had the least effective inhibitory effects across all strains. Notably, water extracts showed the weakest inhibitory effects against all strains.

The current study demonstrated the efficacy of crude extracts of Cenchrus biflorus Roxb. against the tested strains of bacteria and fungi and also discussed their potential application as antibacterial agents for combating infectious diseases. The compositions derived significant antimicrobial properties, making them suitable for patent use in pharmaceutical formulations, nutraceuticals, and natural preservatives.

Endovascular aortic repair involves the placement of stents through minimally invasive methods to seal rupture sites near the aortic inflow tract, thereby preventing blood entry into the false lumen and promoting thrombosis, which reduces the risk of aortic rupture. Endovascular stents typically consist of a metal framework and a flexible membrane graft designed to reopen obstructed aortic segments and maintain blood flow through the true lumen. Consequently, stents are widely used to treat aortic expansion diseases and aortic occlusive stenosis. However, traditional stents have limitations in terms of adaptability to complex anatomical structures, long-term durability, biomechanical stability, and reliance on radial support force for fixation, lacking active fixation mechanisms. These shortcomings remain the primary causes of postoperative complications, significantly impacting the quality of life for patients with aortic dissection.

Integrating patent and academic literature, the research status of the endovascular stent was discussed in depth, and the main factors for the optimal design of the stent (geometry, pattern configuration, additional fixtures, and optimization methods) were analyzed and summarized according to the complications targeted by the repair device.

The composition structure, working principle, and development status of the stent grafts under review are elaborated in detail. Stent grafts attempt to alleviate postoperative complications through three approaches: enhancing the flexibility of the stent framework, improving the fit between the vessel wall and the stent, and reducing vascular injury. Blood flow guiding channels are established to alleviate the obstruction of branch blood flow. Additional self-anchoring devices are added to adapt to the dynamic remodeling of blood vessels.

The effects of various factors, including geometric parameters, structural design, and parameter optimization techniques, on the optimization of stent primary mechanical performance are discussed. The current research status of functional improvement methods for stents is also summarized.

Refining the quantitative relationship between stent structural parameters and mechanical performance, as well as exploring the balance criteria between flexibility and radial support force, represent promising directions for future development. These objectives necessitate further in-depth analysis and research.

Vernonia anthelmintica (L.) Willd., commonly known as Kalijiri, has been used for the treatment of different ailments, including stomach aches, skin diseases, asthma, and cough, and is popular as a powerful anthelmintic agent.

This study aimed to evaluate the pharmacognostical, physicochemical, and phytochemical parameters along with the in-vivo antiulcer activity of the seeds of Vernonia anthelmintica (L.) Willd.

The pharmacognostical evaluation included macroscopic and microscopic characterization of the seed and powder of Vernonia anthelmintica (L.) Willd. Physicochemical parameters, such as moisture content, ash values, and extractive values, were evaluated, and fluorescence analysis was carried out. Phytochemical screening, including total flavonoid content (TFC) and total phenolic content (TPC), was conducted, along with antioxidant analysis. The anti-ulcer activity of ethanolic extract of V. anthelmintica seeds was also evaluated using an aspirin-induced ulcer model by employing multiple biochemical and histopathological assessment parameters.

The morphological characteristics of V. anthelmintica seeds demonstrated that the seed was 0.8-1 cm in length and 0.1-0 in width, with a bitter taste, whereas microscopical findings revealed the presence of pericarp, endosperm, sclerenchymatous zone, parenchymatous zone, bundles of sclereids, seed coat, and prismatic crystals. Moisture content and ash values, including total ash, acid-insoluble ash, and water-soluble ash, were approximately 10.05%, 8.50%, 2.06%, and 4.35%, respectively. Extractive values of different solvents (petroleum ether, chloroform, benzene, ethanol, and water) were approximately 16.00%, 15.80%, 2.00%, 7.05%, and 1.06%, respectively. Moreover, fluorescence analysis revealed a characteristic brown colour. Preliminary phytochemical analysis showed the presence of phenolic, carbohydrates, proteins, flavonoids, saponins, diterpenes, steroids, and amino acids. The TFC and TPC revealed that the ethanolic extract contained more phenolic content, whereas the aqueous extract contained more flavonoids. The ethanolic extract exhibited 90% DPPH radical scavenging activity at a concentration of 100 µg/mL, while the aqueous extract showed 85.71% activity at the same concentration. In comparison, ascorbic acid demonstrated 94.32% scavenging activity at 40 µg/mL. The ethanolic extract of V. anthelmintica seeds at doses 200 and 400mg/kg showed a significant decrease in the ulcer index values, gastric volume, and total acidity levels, whereas an increase was observed in the SOD and GSH levels. The extract demonstrated a moderate effect on the levels of Hb and total protein when compared with the disease control group. The histopathological findings revealed the antiulcer potential of ethanolic extract of V. anthelmintica seeds at both doses.

This study confirmed the identity, quality, and bioactive content of V. anthelmintica seeds, highlighting strong antioxidant and significant antiulcer activity of the ethanolic extract. The results support traditional use and suggest potential for developing patented herbal formulations, encouraging further research on its therapeutic applications.

This research plays a crucial role in raising awareness about the gastroprotective potential of V. anthelmintica. It encourages researchers to explore and further investigate its gastroprotective properties across varying doses and alternative screening models other than those used in this study.

In this study, the coelomic fluid of Eisenia fetida/andrei species was used for the first time to prepare an anti-aging serum, and its antioxidant and antibacterial properties were investigated. In addition, its cytotoxicity on mouse fibroblast cells was measured as material for the production of natural anti-aging products.

This study investigates the antibacterial, antioxidant, and cytotoxic properties of coelomic fluid extracted from Eisenia fetida/andrei. Earthworms were cultured for a year, and their coelomic fluid was extracted using an electroshock method, sterilized, and lyophilized into powder. Antibacterial activity was tested against Escherichia coli and Staphylococcus aureus using MIC assays. Antioxidant properties were evaluated using the DPPH radical scavenging assay. Cytotoxicity effects on L929 and NHEK cell lines were assessed using MTT assays. Oxidative stress and enzymatic activities were analyzed by measuring malondialdehyde (MDA) levels and catalase activity in NHEK cells treated with coelomic fluid. A serum formulation incorporating coelomic fluid was prepared and subjected to stability tests, including pH, temperature, mechanical, and heavy metal residue analysis. Antibacterial and antioxidant properties of the serum were also evaluated. Statistical analyses were conducted using SPSS software (version 0.26). Results highlight the multifunctional potential of coelomic fluid for biomedical and cosmetic applications.

Coelomic fluid exhibited antibacterial activity with MICs of 0.15 mg/mL for both E. coli and S. aureus, showing significant inhibition at higher concentrations. Ciprofloxacin and penicillin demonstrated stronger effects compared to the coelomic fluid. Antioxidant activity increased with concentration, achieving 77% inhibition at 10 mg/mL, with an IC50 of 10.67 mg/mL. Cytotoxicity analysis revealed no significant toxicity below 20 mg/mL, with enhanced cell viability at 2.5–5 mg/mL and restorative effects on fibroblasts at 10 mg/mL. Oxidative stress assays indicated reduced lipid peroxidation and increased catalase activity without inducing significant oxidative stress. Measurement of residues of mercury and lead in the sera showed that they were less than 0.01 ppm for mercury and less than 0.03 and 0.05 ppm for lead, respectively. These levels are below the U.S. Food and Drug Administration's approved limits for these metals. Aqueous serum containing coelomic fluid showed similar antibacterial and antioxidant properties, emphasizing its potential for cosmetic and pharmaceutical applications.

These results show that the use of earthworm coelomic fluid in skin care serum slows the aging process and restores damaged cells. The results of the present study can be considered as a patent.