Avicenna journal of medical biotechnology最新文献

Background: The enzyme L-lysine-α-oxidase from Trichoderma harzianum Rifai is a promising anticancer, antifungal and antibacterial agent. Intensive exploring of its physico-chemical properties and possible ways of application requires sufficient amounts of the protein which in turn depends on good techniques of cultivation of the microorganism producer, enzyme soft isolation and purification, and storage.

Methods: An improved method has been suggested for isolation and purification of the enzyme. A specific combination of column sorbents was adapted and gradient elution with sodium chloride was applied to elevate the yield of the enzyme. The inductive influence of Metabolic Products (MP) of the Brevibacterium species, along with fungal metabolites of Ulocladium sp. and Trichoderma sp. was tested. The enzyme activity assay was based on the detection of oxidized dimethylbenzidine in a peroxidase reaction coupled with an L-lysine-α-oxidase reaction. Some enzyme properties were additionally explored.

Results: The upgraded technique of isolation and purification resulted in a yield of enzyme of about 79%. All strains of Brevibacterium sp. proved to be potent enhancers of L-lysine-α-oxidase activity and concomitant activities. The induced enzyme appeared to be less specific but more thermostable. Possible application scopes for the enzyme with modified properties are discussed. Phosphate buffer solution (pH=5.6) appeared to be the best one for long-term storage of the enzyme.

Conclusion: A significant inducing effect of MP of Brevibacterium sp. on L-lysine-α-oxidase has been detected, and its isolation and purification techniques have been improved.

Background: This study addresses the increasing prevalence of type 2 Diabetes Mellitus (T2DM) and the need for alternative, cost-effective medications. The research problem focuses on the need to further study the dose effectiveness of Momordica charantia Extracts (MCE) on T2DM parameters, including Glucagon-Like Peptide 1 (GLP-1), Dipeptidyl Peptidase 4 (DPP-4), alpha glucosidase, glucosetransporter 5 (GLUT5), and pancreatic tissue histopathology.

Methods: The methodology employed an experimental research design with 30 Wistar rats divided into six groups, each receiving different inductions and doses. Parameters were measured using Elisa, and histological analysis of pancreatic tissue was conducted using HE staining.

Results: The Kruskal-Wallis test revealed significant differences in each group for the GLP-1 (p=0.003). However, the DPP4 test suggested a lack of significant difference in each group (p=0.192), and the GLUT5 test showed insignificant changes between each group (p=0.119). The ANOVA analysis on alpha-glucosidase revealed no statistically significant differences among the groups (p=0.202). Additionally, a qualitative examination of the histological analysis of pancreatic tissue indicated an improvement in the condition of the pancreatic tissue.

Conclusion: MCE can increase GLP-1 levels, lower DPP-4, lower alpha-glucosidase, and raise GLUT5. However, there are no significant differences between other groups and the morphology of pancreatic tissue in rat model T2DM at a dose of 300 mg/kg.

Background: The aim of the present study was to investigate the potential of Nanochelating-based copper to accelerate the wound healing process and prevent infection in burn wounds.

Methods: Six to eight-week- old female BALB/c mice were burned with a 1 cm² heated copper plate on the left flank and then divided into four treatment groups, treated with C8 (nanochelating-based CuNPs), cold cream (supplementary materials) as a control drug, Silver Sulfadiazine and no treatment, respectively. Skin tissue samples were taken from the mice on days 0, 3, 8, 15 and 24. One piece was fixed in 10% neutral buffered formalin for pathological examination and the others were stored at -80°C until used for pro-inflammatory and growth factor gene expression.

Results: The healing process in the group treated with 10 mg/ml C8 was significantly faster, and the survival rate of the mice in this group was significantly higher than in the other groups. The pro-inflammatory genes were expressed and down-regulated earlier in the C8 treated mice. Histopathology confirmed the higher cure rate in the group treated with 10 mg/ml C8 compared to other control groups.

Conclusion: C8 has beneficial effects on the healing of burn wounds and the effective dose of this compound should be further investigated. The present study demonstrates the anti-inflammatory properties of nano-chelate-based copper particles' on mouse skin burns. This research opens up new possibilities in dermatology and burn therapy and highlights the potential of copper-based formulations in the treatment of burn injuries.

Background: Since the initial outbreak, the SARS-CoV-2 virus has continued to circulate and mutate, resulting in the emergence of new viral sublineages. Due to the lack of effective protection and therapeutic measures against these new variants, the virus is able to further evolve and diversify. This study aimed to screen a phage antibody library to identify monoclonal antibodies in single-chain variable fragment (scFv) format that target the Receptor Binding Domain (RBD) of different SARS-CoV-2 strains. The newly discovered scFv has the potential for use as a diagnostic or therapeutic option against SARS-CoV-2.

Methods: The RBD protein was produced, purified, and used as an antigen during biopanning. Six rounds of panning enriched RBD-specific phages and the binding affinity of binders were monitored by polyclonal phage ELISA. Subsequently, monoclonal phage ELISA was employed to identify specific binders. After sequence confirmation, the reactivity of the isolated anti-RBD scFv was evaluated. Additionally, bioinformatics tools determined the interaction between selected scFv and SARS-CoV-2 strains.

Results: The ELISA analysis demonstrated that the expressed RBD retains its structural integrity and effectively interacts with antibodies present in the sera of COVID-19 patients. Through screening a phage display library, a strong-binding scFv for RBD was discovered, which can effectively neutralize SARS-CoV-2 and its novel variants.

Conclusion: The findings of this study have led to the discovery of a novel scFv that effectively neutralizes SARS-CoV-2 strains, offering immense potential for research and therapy purposes.

Background: The utilization of biomarkers is a way to assess the efficacy of recently created anticancer drugs. MiRNAs, telomerase, and Bcl-2 are extensively utilized as biomarkers for this purpose. This study aims to evaluate the comparison of the newly synthesized platinum compounds such as Platinum Azidothymidine (Pt-AZT) with Platinum Levetiracetam (Pt-Lev) on HepG2 cancer cell lines via the biomarkers.

Methods: In this study, cells were divided into four groups: Group A (HDF cells) were the normal negative control group, group B were HepG2 untreated cancer cells, and groups C and D were treated cancer cells with Pt-AZT and Pt-Lev, respectively. After evaluating the LC₅₀ for the drugs by MTT test, the relative gene expression of the biomarkers was determined by qPCR.

Results: The results showed a significant decrease for antiapoptotic genes including miRNA-21 (5.1±0.014), telomerase (0.56±0.48), Bcl-2 (0.41±0.276) in group D, whereas in group C was more than group D for miRNA-21 (6.0±0.141), telomerase (3.49±0.231), Bcl-2 (4.93±0.276) also there was a significant increase in miRNA-122 (33.97±0.04) in group D, whereas in group C was (28.36±0.007) and so lower than group D. Most of the investigated groups showed a significant difference (p<0.05). In addition, there were widespread apoptotic regions in Pt-Lev treatment compared to Pt-AZT.

Conclusion: The advantages of using Pt-Lev were more powerful anticancer effects on biomarkers through inhibition of antiapoptotic and stimulation proapoptotic factors and also lower side effects and lower drug resistance than Pt-AZT; therefore, it can be considered a more effective anti-cancer therapy.

Background: Citrus limon seeds have limited exploration, compared to other parts. The increasing prevalence of mosquito menace prompted the investigation into the repellent effects of the seed n-hexane extract and fractions of Citrus limon (Rutaceae) against adult Anopheles gambiae sl mosquitoes. The cytotoxicity and chemical constituents of the 100% n-Hexane Fraction (HF) were also evaluated.

Methods: Successive extraction was done with cold-maceration using, n-hexane, Ethyl Acetate (EA) and methanol. The three solvent extracts were screened against Anopheles gambiae sl mosquitos, using human bait method, at doses, 1.5, 2.5 and 5.0 mg/ml and with Deet and acetone used as positive and negative controls respectively. The n-hexane extract was further fractionated with column chromatography. Ten fractions (100% nhexane, 10, 20, 30, 40, 60, 80% EA in n-hexane, 100% EA, 50% methanol in EA, and 100% methanol) were collected, and all tested at a single optimized dose of 5 mg/ml for 2 hr. The cytotoxicity and the chemical constituents of HF were determined using brine shrimp lethality test and gas chromatography-mass spectrometry respectively.

Results: N-hexane extract and HF, both at 5 mg/ml, showed the most protective effect of 97.52 and 89.40% respectively, compared with standard (Odomos Deet) 100% activity, with no significant difference (p>0.05) between the three. HF was toxic with LC₅₀ values of 29.2354 ppm. Thirty compounds representing 79.25% of HF were identified. Major compounds present in the fraction were heptadecanamine (8.31%), N-(3-methylbutyl) acetamide (7.89%), 2-ethyl- 1-hexanamine (6.65%).

Conclusion: This promising repellent activity further justified the traditional usage of Citrus limon as mosquito repellent.

Background: Hand, Foot, and Mouth disease is an acute infectious disease caused by a group of enteroviruses, including Coxsackievirus A16 and Enterovirus 71. EV-A71-causing disease can give rise to severe complications in children, leading to meningitis, encephalitis, and even death. A potential approach to prevent the virus spread is inhibiting the invasion of EV-A71 into target cells, thereby helping to prevent not only the spread of EV-A71 in the community but also lessen the risk of outbreaks. EV-A71 cell's receptor, human scavenger receptor class B member 2, SCARB2, was used as a trap to gather the virus and limit its spreading.

Methods: In this study, the recombinant receptor was expressed using Escherichia coli (E. coli) system. SCARB2 proteins expressed from E. coli BL21(DE3), and E. coli SHuffle^® T7 Express were in inclusion bodies and subsequently refolded into soluble forms with recovery efficiencies of 57.57, and 82.2%, respectively. The presence of intramolecular disulfide bridges in the refolded SCARB2 was examined by SDS-PAGE in combination with Dithiothreitol (DTT). The two proteins were then utilized to evaluate the interaction with EV-A71 capsid by Indirect Enzyme-Linked Immunosorbent Assay (ELISA) at different pH levels to compare the adhesion efficiency.

Results: The results showed that SCARB2 protein expressed from E. coli SHuffle^® T7 Express with disulfide bond modifications had better adhesion to the viral capsid. Notably, when the medium pH was lowered to acidic levels, the binding efficiency of recombinant receptors to the viral capsid increased.

Conclusion: To our knowledge, this study reported for the first time the activity of SCARB2 under extreme pH conditions and also revealed the crucial role of disulfide bridges in the interaction with EV-A71's capsid. This finding contributed to the strategy using recombinant SCARB2 as a viral trap.

The integration of Artificial intelligence (AI) in biotechnology presents significant ethical challenges that must be addressed to ensure responsible innovations. Key concerns include data privacy and security, as AI systems often handle sensitive genetic and health information, necessitating robust regulations to protect individuals' rights and maintain public trust. Algorithmic bias poses another critical issue; AI can reflect existing biases in training data, leading to inequitable healthcare outcomes. Transparency in AI decision-making is essential, as "black box" models hinder trust, especially in drug discovery and genetics. Ethical implications of genetic manipulation require careful scrutiny to define the limits of human intervention. Additionally, societal impacts must be considered to ensure equitable distribution of AI benefits, preventing the exacerbation of disparities. Engaging diverse stakeholders, including ethicists and policymakers, is vital in aligning these technologies with societal values. Ultimately, prioritizing ethics will allow us to harness AI and biotechnology's potential while safeguarding human rights and promoting equity.

Background: Stress during pregnancy significantly impacts offspring early physiological programming. Herbal remedies are frequently used by pregnant women to enhance their wellbeing. Moringa oleifera Leaf Extract (MoLE) is believed to have both anti-stress and antioxidant properties which can act as a Selective Estrogen Receptor Modulator (SERM) that regulate activities of estrogen, and can have different effects on different tissues. Goal of this study is to compile information on molecular docking analysis of phytochemicals found in MoLE targeting Estrogen Receptor-alpha (ER-α) and assess effects of MoLE administration on dam's and fetal brain tissues and placenta, during gestational stress.

Methods: Phytochemical study of MoLE was determined using Gas Chromatography-Mass Spectrometry. Molecular docking technique was employed to predict aspects of interaction and binding affinities energy of bioactive phytocompounds in protein site of ER-α using autodock tools. 30 apparently healthy pregnant Albino-Wistar rats were randomly placed into 6 groups of 5 rats per group and exposed to Chronic Unpredictable Stress (CUS) protocol for two weeks, as follows: Group I (water and normal rat chow ad libitum), Group II (CUS protocol only), Group III (5 mg/kg body weight/day of MoLE), Group IV (10 mg/kg body weight/day of MoLE), Group V (CUS protocol +5 mg/kg body weight/day of MoLE), Group VI (CUS protocol +10 mg/kg body weight/day of MoLE).

Results: This study found that 1-Propanol, 3,3'-oxy bis- and 1, 2, 3-Trimethyldiaziridine are most potent ligands for ER-α among all 41 compounds. Photomicrograph examination of tissues from stressed rats showed mild to severe alterations in histology. Consumption of MoLE during chronic stress showed mild to moderate protective effects.

Conclusion: These findings suggest that 1-Propanol, 3,3'-oxy bis- and 1, 2, 3-Trimethyldiaziridine can be further investigated for development of novel therapeutics.