Recent Patents on Biotechnology - Online First

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.

Aeromonas hydrophila, a rod-shaped, gram-negative bacterium, is frequently found in aquatic surroundings and additionally present in drinking water, sewage, and food sources. This microbe is gaining recognition as a potential threat to health, classifying it as an emerging pathogen. Biosurfactants are microbial-derived compounds that share hydrophilic and hydrophobic moieties that are surface active. This study aimed to investigate the effect of biosurfactant isolated from Actinobacteria on the expression of the bfp gene of A. hydrophila isolated from children's stool samples to 
patent the ideal method in Qom, Iran, from May 2022 to March 2023.

Actinobacteria were isolated from soil samples of the desert areas of Qom province, Iran. Biochemical and molecular tests of 16S rRNA were used to identify Actinobacteria isolates. The produced biosurfactant was investigated by methods of hemolysis, oil droplet destruction, lipase production, oil expansion, emulsifying activity, and surface tension reduction measurement. The structure of biosurfactant was investigated by Fourier transform infrared spectroscopy (FTIR) analysis, and its effect on bfp gene expression was measured. Also, isolates of A. hydrophila were obtained from stool samples of children referred to Hazrat Masoumeh Hospital in Qom from May 2022 to March 2023. Then, the effect of a biosurfactant isolated from Actinobacteria on the bfp gene expression of A. hydrophila isolates was measured by RT-PCR.

Based on sequencing data, the Streptomyces genus with the ability to produce biosurfactant was isolated from the soil of the studied area, which could reduce the expression of the bfp gene after treatment with biosurfactant in clinical isolates of A. hydrophila.

The biosurfactant-producing isolates were identified as Streptomyces spp. The results indicated that the biosurfactant significantly decreased bfp gene expression in A. hydrophila. This emphasizes the potential of biosurfactants to eliminate microorganisms by reducing virulence gene expression, inhibiting biofilm formation, demonstrating antimicrobial activity, and improving emulsification. The study supports the idea that biosurfactants can interfere with bacterial mechanisms that cause disease, such as biofilm formation, which is critical for pathogen persistence and resistance. Previous research also confirms the antipathogenic activity of natural isolates against A. hydrophila.

The findings of the present study show that the desert soils of Qom province are a potential area for finding actinobacterial isolates with the ability to produce biosurfactants and influence the expression of pathogenic genes of clinical isolates of A. hydrophila.

This study aims to develop an efficient and reproducible in vitro protocol for high-frequency embryogenic callus induction and subsequent plant regeneration in multiple sorghum (Sorghum bicolor L. Moench) cultivars, thereby establishing a foundation for genetic transformation, mutation breeding, and other biotechnological applications aimed at enhancing sorghum crop improvement and productivity.

Sorghum (Sorghum bicolor (L.) Moench) is an important cereal crop known for its adaptability to harsh environments and nutritional value. Despite its significance, sorghum remains challenging for in vitro propagation due to difficulties in regenerating callus tissue, especially from monocotyledonous explants. Callus induction and regeneration protocols are crucial for genetic transformation, mutation breeding, and biotechnological applications in sorghum improvement.

To establish an effective in vitro protocol for callus induction and subsequent plant regeneration using different sorghum cultivars, optimizing conditions for high-frequency embryogenic callus formation and plant regeneration.

Six sorghum cultivars (IS 3477, IS 33095, IS 7155, IS 2898, IS 7005, and IS 1202) were selected. Immature inflorescence explants were cultured on a modified Murashige and Skoog's (MS) medium with 3% sucrose, 0.8% agar, and 2.0 mg/l 2,4-D for callus induction. After 14 days, embryogenic and non-embryogenic calli were distinguished. Regeneration media were optimized using embryogenic calli, with 1.5 mg/l 6-benzylaminopurine (BAP) for shoot development and 1 mg/l NAA (1-naphthaleneacetic acid) in a half-strength MS medium for root development.

Two distinct forms of calli were observed: a non-embryogenic light yellow callus and a white, granular embryogenic callus. Embryogenic callus induction frequency varied from 40% to 96% among the cultivars, with IS 3477 and IS 33095 exhibiting the highest frequencies (96% and 88%, respectively), while IS 1202 showed the lowest (40%). Regenerated shoots were successfully developed within 6-18 days and later transferred to a rooting medium, resulting in healthy plantlets. Transplanted plantlets showed normal growth and no morphological abnormalities in the field.

This study provides a reliable protocol for efficient callus induction and plant regeneration in multiple sorghum cultivars. The optimized conditions can be utilized for genetic studies, crop improvement, and biotechnological applications, thus contributing to the advancement of sorghum breeding and biotechnology research.

The increasing prevalence of Staphylococcus haemolyticus infections in community and hospital settings presents a significant health challenge due to growing antibiotic resistance and biofilm formation.

This study aims to:(1) perform a molecular analysis of prevalent native strains in Anbar, Iraq, (2) differentiate between various pathogenic strains using multilocus sequence typing (MLST) to enhance epidemiological and surveillance efforts by relevant patents on molecular diagnostics and pathogen typing. The objective is to trace the origins of these strains and distinguish between invasive and indigenous strains. While S. haemolyticus is generally part of the normal human microbiota, it can lead to serious infections in individuals with prior injuries or surgical procedures. It is particularly skilled at developing antibiotic resistance, making it a leading cause of hospital-acquired infections, largely through the staphylococcal cassette chromosome mec (SCCmec). Methicillin-resistant S. haemolyticus (MRSH) has developed resistance to oxacillin/cefoxitin through SCCmec acquisition, and hospital-associated MRSH strains are increasingly resistant to multiple antibiotics.

The preparation of blood agar medium followed the manufacturer's guidelines. After autoclaving at 121ºC for 15 minutes, the medium was cooled to 50ºC. The mixture was then thoroughly mixed and poured into sterile Petri dishes. This medium is used for isolating and cultivating bacteria, as well as for detecting hemolytic activity and identifying the type of hemolysis. Genomic extraction and molecular screening of multidrug-resistant (MDR) isolates were performed, followed by MLST analysis. Data were processed using the University of Nebraska Medical Center's pubMLST website.

To explore the genetic relationships among S. haemolyticus strains, their genomic DNA was analyzed using MLST typing based on the protocol from the MLST Institute database. All S. haemolyticus isolates in the study underwent MLST gene screening through PCR to verify the presence of housekeeping genes (arc, SH1200, hemH, leuB, SH1341, cfxE, and ribose ABC). PCR electrophoresis results demonstrated successful amplification of all target genes, confirming their appropriateness for MLST analysis. Three isolates were recognized as novel global strains, designated ST153, ST154, and ST155. In addition, five other strains were previously registered as ST3, ST9, ST29, ST123, and ST124.

The findings diverge from the established global understanding of type distribution in Asia. To combat the spread of highly resistant strains, it is crucial to monitor virulence factors and antibiotic resistance closely.

Bacillus amyloliquefaciens contains several fungal inhibitory compounds, such as peptides and lipopeptides, representing the remarkable potential for biotechnological, agricultural, and biopharmaceutical applications.

This research aimed to extract and characterize iturin A as the key antagonism factor of Bacillus amyloliquefaciens M13RW01 toward pathogenic fungi, using HPLC and mass spectrometry (MS) analysis.

For this study, Bacillus amyloliquefaciens strain M13-RW01 isolated from Isfahan soil was used. The lipopeptide compounds of B.amyloliquefaciens were examined for antagonistic performance against Aspergillus niger PTCC 5010, Mucor hiemalis PTCC 5292, Fusarium oxysporum CBS 62087, and Penicillium chrysogenum PTCC 5037 by well diffusion and percentage of growth inhibition. The crude extract was run on Waters μBondpak C18 column in the HPLC system to separate the antibiotics. Major antibiotics were analyzed based on MS.

HPLC analysis demonstrated that the lipopeptide compound is similar to iturin A. Moreover, MS analysis of these compounds and purified iturin A revealed a high similarity between them, with the same molecular ion peaks identified. Results showed that the produced lipopeptides by Bacillus amyloliquefaciens were of iturin A genum. The molecular ion peaks of the B.amyloliquefaciens M13RW01 methanolic fraction were at 1027.10, 1043.05, 1058, 1066, 1072, 1088.95. These compounds restrained fungal germination and growth. Inhibition growth percentages were 79.28, 76.13, 84.47and 59.15% for Aspergillus niger, Mucor hiemalis, Fusarium oxysporum, and Penicillium chrysogenum, respectively.

According to the present study, B.amyloliquefaciens M13RW01 lipopeptides are able to inhibit the growth of some fungi. B.amyloliquefaciens M13-RW01 isolated from Isfahan soil plays an essential part in antagonizing pathogenic fungi. Thus, this antifungal lipopeptide is supposed to be a biological protection agent for agricultural products and patents.