Biochemistry Research International最新文献

Type 2 diabetes is a complex metabolic disorder characterised by altered amino acid metabolism. This study investigates plasma amino acid profiles and specific aminoacidurias in patients with type 2 diabetes. We recruited 115 patients with type 2 diabetes and 115 healthy controls. Urine amino acids were analysed using paper chromatography, while serum amino acids were analysed using gas chromatography-mass spectrometry (GC-MS). Diabetes-induced alterations in amino acid metabolism are multifaceted. Hyperglycemia and insulin resistance can lead to increased gluconeogenesis, resulting in the depletion of certain amino acids, such as glutamine and arginine. Conversely, the increased availability of branched-chain amino acids (BCAAs) such as leucine, isoleucine and valine can contribute to insulin resistance and impaired glucose metabolism. Significant alterations in plasma and urine amino acid levels were observed in patients with type 2 diabetes, including increased levels of BCAAs and decreased levels of glutamine and arginine. These changes correlated with disease severity and metabolic parameters. Understanding these alterations is crucial for developing novel therapeutic strategies and diagnostic tools for type 2 diabetes management. Our findings provide new insights into amino acid metabolism in type 2 diabetes, identifying potential biomarkers for disease diagnosis and monitoring. These results suggest avenues for developing therapeutic strategies and contribute to understanding the complex relationship between amino acid metabolism and type 2 diabetes. The study highlights the potential of amino acid profiling as a diagnostic and therapeutic tool in type 2 diabetes management.

L-arginine induces the expression of utrophin in skeletal muscle cells, so it has been proposed as a pharmacological treatment to attenuate the symptoms of Duchenne muscular dystrophy (DMD). On the other hand, it has been described that one of the pathways that participates in the expression of utrophin in muscle is the Neuregulin-1 (NRG-1)/ErbB receptors pathway. Several studies have postulated that disintegrin and metalloprotease-17 (ADAM17) causes the proteolytic processing of NRG of transmembrane, allowing the release of NRG to the medium, which when joining its ErbB receptor activates the signaling pathway that triggers utrophin transcription. The aim of this study was to evaluate the effect of L-arginine in the activation of NRG-1/ErbB pathway and utrophin mRNA levels in C2C12 cells, and the participation of ADAM17 in this process. Our results indicate that L-arginine induces phosphorylation of ErbB2 and increases utrophin mRNA levels in C2C12 myotubes, with a maximum increase of 2-fold at 4 h post-stimulation. This effect is not observed when the myotubes are stimulated in the presence of GM6001 (general metalloprotease inhibitor) or PD-158780 (specific inhibitor of ErbB receptor phosphorylation). Experiments performed by flow cytometry suggest that L-arginine stimulates ADAM17 activation in our study model. Furthermore, immunofluorescence analysis supports our findings that L-arginine stimulates ADAM17 increase in treated myotubes. However, our results using pharmacological inhibitors suggest that ADAM17 does not participate in utrophin expression in C2C12 cells treated with L-arginine. The results obtained help to clarify the mechanism of action of L-arginine in the expression of utrophin in muscle cells, which will contribute to the design of new therapeutic strategies in pathologies such as DMD.

The use of local plant extracts in drinking water purification represents a sustainable alternative in the fight against diseases linked to unsafe water consumption in rural areas. The aim of this study was to evaluate the biochemical composition and antimicrobial activity of four local plant extracts used in rural areas of Burkina Faso to purify drinking water: Moringa oleifera seeds, Boscia senegalensis seeds, Opuntia ficus-indica cladodes, and Aloe vera leaves. These four extracts were then subjected to biochemical screening to identify phytocompounds, followed by quantification and evaluation of their antibacterial properties on ten pathogenic bacterial strains. The screening results revealed the presence of a variety of molecules (phenolic compounds, alkaloids, saponosides, etc.) in the different extracts studied. From a quantitative point of view, M. oleifera and B. senegalensis seeds showed a high total protein content (34.5 and 24.6 g/100 g DM). A. vera and O. ficus-indica extracts showed high levels of total carbohydrates (20.4 and 35.52 g/100 g DM) compared with total lipids and proteins. The same applies to phenolic compounds, which were also high in A. vera and O. ficus-indica extracts (17.42 and 26.5 mg GAE/100 mg DM) compared to M. oleifera and B. senegalensis seeds. In terms of antibacterial properties, the four extracts studied showed inhibition diameters ranging from 7.33 to 16.33 mm. These results reflect the ability of the different extracts to eliminate pathogenic microorganisms present in water. Overall, this study stands out for its innovative character, offering an in-depth understanding of the biochemical composition and antimicrobial properties of four distinct extracts of local plants commonly used in traditional drinking water purification practices. It enriches existing knowledge by providing new data on the biochemical composition and bioactivity of these extracts. In particular, the study highlights the synergistic effects of the bioactive compounds present, underlining their essential role in improving the sanitary quality of water consumed in rural areas, where sustainable and accessible solutions are crucial.

Proteins have shown varying degrees of antioxidant activity. This study examined the potential mechanisms of interactions between proteins and radicals using chemical kinetics and computational methods. The study quantified the antioxidant activity of lactate dehydrogenase (LDH) and bovine serum albumin (BSA) through Trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. BSA was about seven times and LDH 12 times more potent as antioxidants for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS^•-) than they were for peroxyl radicals. According to the evaluation of Trolox equivalents (TE) of 20 proteinogenic amino acids, tryptophan (with a TE value of 101 μmol TE/μmol) exhibited the highest antioxidant activity for ABTS^•-, followed by tyrosine (38.7 μmol TE/μmol) and cysteine (30.5 μmol TE/μmol), lysine (0.193 μmol TE/μmol), arginine (0.0325 μmol TE/μmol), valine (0.0280 μmol TE/μmol), histidine (0.00689 μmol TE/μmol), and leucine (0.00560 μmol TE/μmol). The EC50 showed a similar order with a swap between valine and histidine. The antioxidant activity of the amino acids and proteins was temperature dependent. The rate laws, activation energy, and pre-exponential factor A of these reactions provided information on the reaction mechanisms, i.e., a biomolecular elementary step for the reaction of ABTS^•- with amino acids tryptophan, tyrosine, cysteine, or protein LDH, and a more complicated mechanism for BSA. The presence of -NH- or hydroxyl groups on aromatic rings enhanced the antioxidant ability of tryptophan and tyrosine. LDH's antioxidant activity did not affect its enzymatic activity, indicating that the radical reaction likely happened on the protein's surface without significantly altering its conformation. The molecular modeling and visualization showed potential reaction sites on the proteins' accessible tryptophan and tyrosine residues. However, the mere surface exposure of tryptophan and tyrosine does not guarantee their antioxidant activities.

Recently, it has been shown that protein phosphatase 2A (PP2A) dysfunction was common in many cancer types and was mediated by various inactivation mechanisms. Although many research studies observed antitumor effect of propolis extracts in various types of cancer, the mechanism of effect are still obscure. In this study, we investigated the effect of propolis on PPP2R1A expression and its relationship with apoptosis in the SW-620 (colorectal cancer), DU-145 and PC-3 (prostate cancer), and MCF-7 (breast cancer) cell lines, with WI-38 (healthy fibroblast) cells serving as the control. Moreover, we aimed to investigate the impact of propolis on apoptosis by analyzing apoptosis markers such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), APAF-1, and caspases-3, -8, and -9. Propolis samples were extracted, and their phenolic compounds were quantified using LC-MS/MS. The RealTime Cell Analysis System-xCELLigence (RTCA-SP) device and software were employed to assess cell viability and cytotoxicity of the propolis samples. The IC₅₀ values for propolis were determined (298 μg/mL for SW-620, 185.6 μg/mL for DU-145, 250.7 μg/mL for PC - 3, 292.9 μg/mL for MCF-7, and 311.2 μg/mL for WI-38). Subsequently, the effects of propolis on PPP2R1A expression and apoptosis markers (TRAIL, Apaf-1, and caspases-3, -8, and -9) were analyzed. When we compared the healthy cell lines to cancer cell lines, a statistically significant increase in caspase-3 (3.62-fold) and in TRAIL (4.38-fold) was observed in the SW-620 cell line after the application of propolis. In addition, in the PC-3 cell line, a 1.4-fold increase in caspase-8 was observed compared with the healthy cell line, which is also statistically significant. Our findings indicated that propolis increased the PPP2R1A levels and apoptosis markers in cancer cell lines. It has been suggested that high PPP2R1A levels induced by propolis treatment might activate the apoptosis pathway. In this study, the inducible effect of propolis on PPP2R1A levels, identified as a new target for cancer treatment, was demonstrated for the first time. The findings suggest that propolis holds promise as a potential cancer therapy by increasing PPP2R1A levels, a key molecule in cancer treatment.

Members of the Costus genus are the conventional medicinal plants used in the therapeutic management of numerous ailments, especially for their antioxidant and pharmacological activities. The crude extract of Costus spicatus was profiled using high-resolution GC-MS and LC-MS/MS techniques to determine possible bioactive compounds that are vital to the antioxidant activity. A total of 52 and 63 bioactive compounds have been detected in GC-MS chromatograms using different solvents (methanol and ethanol) in C. spicatus leaf extracts, representing the presence of certain bioactive compounds. The identified bioactive compounds in both extracts which exhibit neuroprotective effects have been confirmed through various literature studies. They are cholestan-3-amine, loliolide, stigmasterol and methylprednisolone acetate, succinimide, fumaric acid, beta-tocopherol and gamma-tocopherol. The aqueous extract possessed the highest antioxidant activity for DPPH scavenging activity and lipid peroxidation inhibition assays, whereas the alcoholic aqueous extract showed superior efficacy for hydroxyl radical scavenging activity. In this study, we performed molecular docking and found that four compounds exhibited promising binding affinities with predicted binding sites on alpha-synuclein. Notably, Androsta [17-16-b] furan-5'-imine, 4'-methylene-3-methoxy-N-cyclohexyl- showed the highest docking interaction score of -7.4, indicating a strong binding affinity. These findings, combined with the presence of bioactive components in the crude extract of C. spicatus, suggest that this plant may possess neuroprotective properties, warranting further investigation for potential industrial applications in the development of neuroprotective agents.

Age-related macular degeneration (AMD) is a severe eye disease in people aged 60 years and older. Although anti-VEGF therapies are effective in treating neovascular AMD (NvAMD) in the clinic, up to 60% of patients do not completely respond to the therapies. Recent studies have shown that blood-derived macrophages and their associated proinflammatory cytokines may play important roles in the development of persistent disease and resistance to anti-VEGF therapy. To address this issue, we constructed an antibody-based bispecific fusion protein that can simultaneously inhibit IL-17-induced inflammation and VEGF-mediated neovascularization. As a result, the bispecific fusion protein 17V05 effectively inhibited multiple proinflammatory cytokines and chemokines, as well as laser-induced choroidal neovascularization (CNV). More importantly, 17V05 also exhibited stronger and longer inhibitory effects than conbercept in vivo. Thus, we provide a novel and promising strategy for treating AMD patients who are not sensitive to anti-VEGF therapies.

Alzheimer's disease (AD), a neurological disorder, is one of the major reasons for memory loss in the world. AD is characterized by a sequela of cognitive and functional decline caused by brain cell degeneration. Paeoniflorin is a monoterpenoid glycoside found in plants of the Paeoniaceae family, which are known for their medicinal properties including dementia. In this project, we report actions of paeoniflorin on the two related cholinesterases (ChE): acetylChE (AChE) and butyrylChE (BuChE). Paeoniflorin, in a dose-dependent (maximum inhibition at 1 mg/mL) manner, inhibited both AChE (0.06-1 mg/mL) and BuChE (0.007-1 mg/mL) enzymes with maximum inhibition of AChE enzyme at 90.3 ± 1.4%, while 99.4 ± 0.3% for BuChE enzyme. The EC₅₀ value for the inhibitory effect of the compound against AChE was 0.52 mg/mL (0.18-1.52), while against BuChE was 0.13 mg/mL (0.08-0.21). The observed ani-ChE action was like an effect also mediated by the known ChE blocker physostigmine. Molecular interactions between paeoniflorin and both ChE enzymes were additionally sought via molecular docking and molecular dynamics simulations for 100 ns, that showed paeoniflorin interacted with the active-site gorge of AChE and BuChE via hydrogen bonds and water bridging with the many amino acids of the AChE and BuChE enzymes. This study presents the ChE inhibitory potential of paeoniflorin against both AChE and BuChE enzymes. With this kind of inhibitory activity, the chemical can potentially increase ACh levels and may have use in the treatment of dementia of AD.

Woodfordia uniflora is a medicinal plant used for the treatment of malaria, toothache, and stomach problems. The root parts of the plant are also used for healing liver disorders. Silica gel chromatography separation of CH₂Cl₂/MeOH (1:1) and MeOH extracts of roots of W. uniflora result in the isolation of three compounds, namely, bergenin (1), β-sitosterol (2), and epigallocatechin 3-gallate (3), reported herein for the first time from the plant. The structure of the isolated compounds was elucidated using NMR (1D and 2D) techniques. Disk diffusion and DPPH assay were used to evaluate the antibacterial and antioxidant activities, respectively. Molecular docking was done by the AutoDock Vina 4.2 program. The pharmacokinetics and toxicity profile of compounds were predicted by Swiss ADME and Pro Tox II online servers. GC-MS analysis roots of W. uniflora result in the identification of five compounds, of which palmitic acid (34.9%) was the major constituent. The antibacterial activity result indicated that the oil extract had promising activity against P. aeruginosa, E. coli, S. pyogenes, and S. aureus with IZ of 14.3 ± 0.81, 15.0 ± 0.0, 15.6 ± 0.47, and 17.6 ± 0.47 mm, respectively, at 5 mg/mL, compared to ciprofloxacin (1Z 27-30.0 ± 0.0 mm) at 30 μg/mL. MeOH and CH₂Cl₂/MeOH (1:1) extract showed inhibition against E. coli (IZ of 13.6 ± 0.47 mm) and P. aeruginosa (IZ of 10.0 ± 0.0 mm), respectively, at 200 mg/mL. Bergenin (1) and β-sitosterol (2) also displayed maximum inhibition of E. coil (IZ of 11.6 ± 0.47) and S. aureus (11.0 ± 0.0 mm), respectively, at 5 mg/mL. The antioxidant activity results showed that CH₂Cl₂/MeOH (1:1) and MeOH extracts, bergenin (1), and compound 3 displayed potent scavenging DPPH radical with a percentage of inhibition of 76.8 ± 0.12, 77.8 ± 0.08, 71.4 ± 0.08, and 91.2 ± 0.16, respectively, compared to ascorbic acid (93.2% ± 0.04%) at 100 μg/mL. The molecular docking analysis showed that all compounds (1-3) exhibited minimum binding energy toward PDB ID: 1HD2 (-5.2 to -6.3 kcal/mol), compared to ascorbic acid (-5.6 kcal/mol), and toward PDB ID: 1DNU (-8.0 to -10.7 kcal/mol) receptors, compared to ascorbic acid (-5.7 kcal/mol). Toward the PDB ID: 4FM9 receptor, β-sitosterol (2) and compound 3 exhibited the best binding free energy of -9.1 and -9.8 kcal·mol, respectively, compared to vosaroxin (-7.8 kcal/mol). The drug-likeness analysis result indicated that bergenin (1) and β-sitosterol (2) obeyed four and five criteria of Lipinski's rule, respectively, and are more likely to be administered orally. The in silico toxicity analysis showed none of the compounds would be cytotoxic, mutagenic, or hepatotoxic. The in vitro antioxidant and antib

The clinical data indicate that diverse parameters characterize breast cancer patients in India, including age at presentation, risk factors, outcomes, and behavior. Alarming incidence and mortality rates emphasize the crucial need for early screening measures to combat breast cancer-related deaths effectively. Quantitative proteomic approaches prove pivotal in predicting cancer prognosis, analyzing protein expression patterns tied to disease aggressiveness and metastatic potential, and facilitating conversant therapy selection. Thus, this study was envisioned with the goal of identifying protein markers associated with breast cancer in Indian women, which could potentially be developed as diagnostic tools and therapeutic targets in the future. Applying label-free proteomic quantitation method and statistical analysis, several differentially expressed proteins (DEPs) were identified in the serum of breast cancer patients compared to controls, including SBSN, ANG, PCOLCE, and WFDC3 (upregulated), and PFN1, FLNA, and DSG2 (downregulated). The expression of SBSN was also validated by western blotting. Statistical methods were employed to proteomic expression data, which highlighted the ability of DEPs to distinguish between breast cancer and control samples. Conclusively, this study recognizes prospective biomarkers for breast cancer among Indian women and highlights the requisite of in-depth functional studies to elucidate their precise roles in breast cancer development. We particularly emphasize on SBSN and PFN1, as these proteins were observed to be progressively overexpressed and under expressed, respectively, in breast cancer samples compared to control samples, ranging from early-stage to metastatic cases.