Biochemistry Research International最新文献

This study focuses on the synthesis of silver nanoparticles (AgNPs) using the extract of Aphania senegalensis leaves. The extraction was done using maceration at room temperature in water for 48 h. The synthesized nanoparticles were characterized by IR, XRD, TEM, and SEM. The thermal stability of these nanoparticles was studied by TGA. The zeta potential was used to define the size, charge distribution, and stability of the nanoparticles. Optimization reactions were carried out based on reaction time, pH, and temperature. The nanoparticles obtained from optimal conditions were evaluated on induced inflammation. The determination of the average diameters and geometry of nanoparticles was carried out by XRD by calculating the lattice constants, and they are between 18.11 and 50 nm. The evaluation of anti-inflammatory activity showed that the nanoparticles are 10 times more active than the extract of Aphania senegalensis leaves. Minimum doses of 10 mg/kg orally and 3 mg/kg were obtained for the plant extract, respectively. These results are promising for the possibility of AgNPs to be used for the treatment of inflammation.

Ovarian cancer's asymptomatic nature, high recurrence rate, and resistance to platinum-based chemotherapy highlight the need to find and characterize new diagnostic and therapeutic targets. While prior studies have linked aberrant expression of fibroblast growth factor 8 (FGF8) to various cancer types, its precise role has remained elusive. Recently, we observed that FGF8 silencing reduces the cancer-promoting properties of ovarian cancer cells, and thus, this study aimed to understand how FGF8 regulates the development of ovarian cancer. LC-MS/MS-based quantitative proteomics analysis identified 418 DEPs, and most of them were downregulated in FGF8-silenced ovarian cancer cells. Many of these DEPs are associated with cancer progression and unfavorable prognosis. To decipher the biological significance of DEPs, bioinformatics analyses encompassing gene ontology, pathway analysis, protein-protein interaction networks, and expression analysis of hub genes were carried out. Hub genes identified in the FGF8 protein network were upregulated in ovarian cancer compared to controls and were linked to poor prognosis. Subsequently, the expression of hub genes was correlated with patient survival and regulation of the tumor microenvironment. Conclusively, FGF8 and associated hub genes help in the progression of ovarian cancer, and their overexpression may lead to higher immune infiltration, poor prognosis, and poor survival.

The glyphosate herbicide is a pesticide widely used in the world and can contaminate soil, air, and water. The objective of this work was to evaluate the toxicity of a glyphosate-based herbicide (GBH) in zebrafish (Danio rerio). Fish were exposed to different concentrations of GBH (0, 50, 250, and 500 µg/L) for 96 hours. Brain, liver, and blood were collected for biochemical and genotoxicity analyses, and behavioral tests were performed. The results showed that there was a reduction in the activity of the antioxidant enzymes of catalase (CAT) and glutathione-S-transferase (GST) in the liver at all concentrations and at the highest concentration in the brain. There was also a reduction in lipid peroxidation in the liver at all concentrations of glyphosate. There was an increase in micronuclei in the blood at the 500 µg/L concentration. However, the count of nuclear abnormalities showed no differences from the control. Interleukin-1beta (IL-1β) generation was inhibited at all concentrations in the liver and at the highest concentration in the brain. No significant differences were found in the behavioral test compared to the control. The results showed that acute exposure to GBH promoted an inflammatory event, which reduced the efficiency of antioxidants, thus producing a disturbance in tissues, mainly in the liver, causing immunosuppression and generating genotoxicity.

Ziziphus spina-christi (Rhamnaceae family) is a medicinal plant traditionally used to treat dandruff, wounds, hair loss, diarrhea, mastitis, abdominal pain, and gastrointestinal complications. To support this, the present work aims to study the in vitro antibacterial and antioxidant activities of compound isolates from the roots of Ziziphus spina-christi along with their in silico computational analyses. Compounds were isolated on silica gel column chromatography and an agar disc diffusion and DPPH radical scavenging assays were employed to study the antibacterial and antioxidant activities, respectively. The ADME and toxicity properties of the compounds were evaluated using SwissADME and ProTox-II online Web tools, respectively. Conversely, the in silico molecular docking studies were attained via a Biovia Discovery Studio Visualizer 2021 in combination with the AutoDock Vina software. The silica gel chromatographic separation of the combined CH₂Cl₂ : CH₃OH (1 : 1) and CH₃OH root extracts afforded trimethyl trilinolein (1), stearic acid (2), 13-hydroxyoctadeca-9, 11-dienoic acid (3), β-sitosteryl-3β-glucopyranoside-6'-O-palmitate (4), and stigmasterol (5). Notably, the in vitro antibacterial study revealed the extract and β-sitosteryl-3β-glucopyranoside-6'-O-palmitate (4) with the highest inhibitory activities (15.25 ± 0.35 and 14.25 ± 0.35 mm, respectively) against E. coli compared to ciprofloxacin (21.00 ± 0.35 mm) at 2 mg/mL. The CH₂Cl₂ : CH₃OH (1 : 1) extract (IC₅₀ : 1.51 µg/mL) and β-sitosteryl-3β-glucopyranoside-6'-O-palmitate (4) (IC₅₀ : 5.41 µg/mL) also exhibited auspicious DPPH scavenging activities, followed by stigmasterol (5) (IC₅₀ : 6.88 µg/mL) compared to the ascorbic acid standard (IC₅₀ : 0.46 µg/mL). The molecular docking analyses unveiled the highest binding affinity by β-sitosteryl-3β-glucopyranoside-6'-O-palmitate (4) (-8.0 kcal/mol) against P. aeruginosa PqsA relative to the ciprofloxacin standard (-8.2 kcal/mol). Furthermore, the organ toxicity predictions showed that all the compounds exhibit no hepatotoxicity and cytotoxicity effects and stigmasterol (5) affords drug-likeness protocols. Overall, the combined experimental and computational investigations of this study support the traditional uses of Ziziphus spina-christi for antibacterial and natural antioxidant applications.

Background: CC-chemokine ligand 18 also known as MIP-4 is a chemokine with roles in inflammation and immune responses. It has been shown that MIP-4 is involved in the development of several diseases including lung fibrosis and cancer. How exactly MIP-4 is regulated and exerts its role in lung fibrosis remains unclear. Therefore, in the present study, we examined how MIP-4 is regulated and whether it acts via its potential receptor Nir-1.

Materials and methods: A549 cells were grown and maintained in DMEM : F12 (1 : 1) and supplemented with 10% FBS and 1000 U of penicillin/streptomycin and maintained as recommended by the manufacturer (ATCC). Cell migration and invasion, immunohistochemistry (IHC), Western blot, qPCR, and siRNA Nir-1 were used to determine MIP-4 regulation and its role in cell migration.

Results: Cell migration was increased following stimulation of cells with recombinant (r) MIP-4 and bleomycin (BLM), whereas quenching rMIP-4 with its antibody (Ab) or addition of the Ab to BLM or H₂O₂ diminished rMIP-4-induced cell migration. Along with cell migration, rMIP-4, BLM, and H₂O₂ induced the formation of actin filaments dynamic structures whereas costimulation with MIP-4 Ab limited BLM- and H₂O₂-induced effects. MIP-4 mRNA and protein were increased by BLM and H₂O₂, and the addition of its Ab significantly reduced treatments effect. Experiments with siRNA investigating whether Nir-1 is a potential MIR-4 receptor indicated that the inhibition of Nir-1 decreased cell migration/invasion but did not totally inhibit rMIP-4-induced cell migration.

Conclusion: Therefore, our data indicate that MIP-4 is regulated by BLM and H₂O₂ and costimulation with its Ab limits the effects on MIP-4 and that the Nir-1 receptor partially mediates MIP-4's effects on increased cell migration. These data also evidenced that MIP-4 is regulated by fibrotic and oxidative stimuli and that quenching MIP-4 with its Ab or therapeutically targeting the Nir-1 receptor may partially limit MIP-4 effects under fibrotic or oxidative stimulation.

The plant Hydnora johannis has been utilized in folk medicine. Analyzing phytochemical composition of dichloromethane/methanol (1 : 1) root part of Hydnora johannis gave oleic acid (1), caffeic acid-2-hydroxynonylester (2), catechin (3), and a pregnane derivative (4). NMR spectroscopy was used to characterize compounds 1-3, while compound 4 was identified through GC-MS analysis and literature comparison. The cytotoxicity of extracts from roots of H. johannis was conducted against MCF-7 cell lines (human breast cancer) by MTT assay. According to the cytotoxicity study, n-hexane extract exhibited a high level of toxicity with 28.9 ± 5.6% cell viability. Antibacterial activity was tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pyogen. The highest bacterial growth mean inhibition zone was measured for catechin (3) (13.72 ± 0.05 mm)) against P. aeruginosa at 0.25 mg/mL and acceptable related to standard. Antioxidant activity was studied by the DPPH assay. Based on the data from the antioxidant study, DCM/MeOH extract (70.32%) and catechin (3) showed good antioxidant activity (65.61%) (IC₅₀ 0.25 μg/mL) relative to that of the positive control (78.21%, IC₅₀ 0.014 μg/mL) at 12.5 μg/mL. In each docking pose, catechin (3) scored higher binding affinity of -7.9, -7.2, and -6.4 kcal/mol towards PqsA, DNA gyraseB, and S. aureus PK, respectively, compared to amoxicillin (-8.1, -6.1, and -6.4 kcal/mol). All five Lipinski rules were obeyed by compounds 1-3, which showed an acceptable drug resemblance. The lipophilicity was computed as less than five (1.47-4.01) indicating a lipophilic property. Catechin (3) obeys Veber's rule implying its good oral bioavailability. Binding affinity scores of catechin (3)-protein interactions are in line with those from in vitro tests, indicating its potential antibacterial effect. The obtained cytotoxicity and antibacterial activity results support the utilization of H. johannis in folk medicine.

Eg5 is a protein encoded by KIF11 gene and is primarily involved in correct mitotic cell division. It is also involved in nonmitotic processes such as polypeptide synthesis, protein transport, and angiogenesis. The scientific literature sheds light on the ubiquitous functions of KIF11 and its involvement in the onset and progression of different pathologies. This review focuses attention on two main points: (1) the correlation between Eg5 and cancer and (2) the involvement of Eg5 in noncancerous conditions. Regarding the first point, several tumors revealed an overexpression of this kinesin, thus pushing to look for new Eg5 inhibitors for clinical practice. In addition, the evaluation of Eg5 expression represents a crucial step, as its overexpression could predict a poor prognosis for cancer patients. Referring to the second point, in specific pathological conditions, the reduced activity of Eg5 can be one of the causes of pathological onset. This is the case of Alzheimer's disease (AD), in which Aβ and Tau work as Eg5 inhibitors, or in acquired immune deficiency syndrome (AIDS), in which Tat-mediated Eg5 determines the loss of CD⁴⁺ T-lymphocytes. Reduced Eg5 activity, due to mutations of KIF11 gene, is also responsible for pathological conditions such as microcephaly with or without chorioretinopathy, lymphedema, or intellectual disability (MCLRI) and familial exudative vitreous retinopathy (FEVR). In conclusion, this review highlights the double impact that overexpression or loss of function of Eg5 could have in the onset and progression of different pathological situations. This emphasizes, on one hand, a possible role of Eg5 as a potential biomarker and new target in cancer and, on the other hand, the promotion of Eg5 expression/activity as a new therapeutic strategy in different noncancerous diseases.

Micromeria graeca L. is a dense chemical source of bioactive compounds such as phenolic compounds, which have various health-related properties. The current study aimed to investigate the impact of different extractor solvents on phenol and flavonoid contents, as well as the antioxidant and antifungal activities of different extracts. Initially, three extractor solvents (methanol, ethyl acetate, and water) were used to prepare the Soxhlet extracts, which were then examined for their polyphenolic content, flavonoid content, and antioxidant potential using three complementary assays (DPPH, FRAP, and TAC). The antifungal capacity against the two fungal strains (Candida albicans and Aspergillus niger) was performed using the method of diffusion on disc. The dosage of phytochemical compounds revealed that the highest values were established in water extract with values of 360 ± 22.1 mg GAE/g dry weight plant and 81.3 ± 21.2 mg RE/g dry weight plant for TPC and TFC, respectively. In addition, the strongest antioxidant activity measured by DPPH and FRAP assays was established in water extract with IC₅₀ values of 0.33 ± 0.23 and 0.23 ± 0.12 mg/mL, respectively, while the methanol extract showed the best antioxidant activity as measured by TAC with an IC₅₀ of 483 ± 17.6 mg GAEq/g dry weight plant. The water extract recorded the most important antifungal activity against Candida albicans with an inhibition zone of 16 ± 1.6 mm and MFC = 500 μg/mL, whereas ethyl acetate extract showed the lowest activity against both studied fungi strains. Micromeria graeca L. contains considerable amounts of bioactive contents with high antioxidant and antifungal potentials, which may make it a promising source of antioxidants and natural antifungal agents.

Bioluminescence has been a fascinating natural phenomenon of light emission from living creatures. It happens when the enzyme luciferase facilitates the oxidation of luciferin, resulting in the creation of an excited-state species that emits light. Although there are many bioluminescent systems, few have been identified. D-luciferin-dependent systems, coelenterazine-dependent systems, Cypridina luciferin-based systems, tetrapyrrole-based luciferins, bacterial bioluminescent systems, and fungal bioluminescent systems are natural bioluminescent systems. Since different bioluminescence systems, such as various combinations of luciferin-luciferase pair reactions, have different light emission wavelengths, they benefit industrial applications such as drug discovery, protein-protein interactions, in vivo imaging in small animals, and controlling neurons. Due to the expression of luciferase and easy permeation of luciferin into most cells and tissues, bioluminescence assays are applied nowadays with modern technologies in most cell and tissue types. It is a versatile technique in a variety of biomedical research. Furthermore, there are some investigated blue-sky research projects, such as bioluminescent plants and lamps. This review article is mainly based on the theory of diverse bioluminescence systems and their past, present, and future applications.

Bacterial and mammalian cells are rich in putrescine, spermidine, and spermine. Polyamines are required for optimum fitness, but the biological function of these small aliphatic compounds has only been partially revealed. Known functions of polyamines include interaction with nucleic acids that alters gene expression and with proteins that modulate activity. Although polyamines can be incorporated into proteins, very few naturally occurring polyaminated proteins have been identified, which is due in part to the difficulty in detecting these adducts. In the current study, bovine albumin and the recombinant universal stress protein from Francisella tularensis were used as models for mass spectrometry analysis of polyaminated proteins. The proteins were covalently bound to putrescine, spermidine, or spermine by the action of carbodiimide or microbial transglutaminase. Tryptic peptides, subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS), were identified using Protein Prospector software. We describe the search parameters for identifying polyaminated peptides and show MS/MS spectra for adducts with putrescine, spermidine, and spermine. Manual evaluation led us to recognize signature ions for polyamine adducts on aspartate, glutamate, and glutamine, as well as neutral loss from putrescine, spermidine, and spermine during the fragmentation process. Mechanisms for the formation of signature ions and neutral loss are presented. Manual evaluation identified a false-positive adduct that had formed during trypsinolysis and resulted in peptide sequence rearrangement. Another false positive initially appeared to be a 71 kDa putrescine adduct on a cysteine residue. However, it was an acrylamide adduct on cysteine for a sample extracted from a polyacrylamide gel. The information presented in this report provides guidance and serves as a model for identifying naturally occurring polyaminated proteins.