Biochemistry Research International最新文献

This study aimed to compare the in vitro biological activities of aqueous leaf extracts from Spinacia oleracea and Basella rubra. Extraction was performed using the maceration method, yielding 25% and 14%, respectively. Phytochemical analysis revealed that S. oleracea had higher total phenolic content (6.2 ± 0.4 mg GAE/g) and flavonoid content (3.8 ± 0.3 mg CE/g) compared to B. rubra. Conversely, B. rubra exhibited stronger antioxidant activity with 79.6% DPPH inhibition versus 64.2% in S. oleracea. In the α-amylase inhibition assay, B. rubra again showed higher enzyme inhibition (72.3%) compared to S. oleracea (51.1%), indicating greater antidiabetic potential. Both extracts displayed no antibacterial activity against E. coli and S. aureus. Hemolysis assays indicated low cytotoxicity: 8.1% for S. oleracea and 5.4% for B. rubra. FTIR analysis identified bioactive functional groups, including phenols, carboxylic acids, glycosides, esters, and alkanes. These findings suggest that both plants possess antioxidant and enzyme-inhibitory potential, warranting further in vivo and phytochemical investigations.

Mushrooms are a source of nutrients and bioactive substances. This work, carried out for the first time, aims to evaluate in vitro and in vivo toxicity and antioxidant potential of the wild edible mushroom from the province of Taza, Morocco: Morchella esculenta (L.) Pers., the extraction is carried out using two methods: aqueous and organic. We assessed the toxicity in vitro on Artemia salina larvae and in vivo on Swiss Albino mice. Antioxidant activity was assessed by H₂O₂, ABTS, FRAP and PR tests. Principal component analysis enabled us to visualise the correlations between the chemical composition, antioxidant activity and in vitro toxicity. The in vitro toxicity results showed that the infused extract was nontoxic (LD50 = 3443.33 ± 20.66 μg/mL) and the diethyl ether macerated was toxic (LD50 = 68.91 ± 2.76 μg/mL). In vivo, this mushroom is not acutely toxic (LD50 ≥ 5000 mg/kg body weight). This study revealed that the decocted was the most active via the H₂O₂ test (23.69 ± 0.61%) and the aqueous macerated was the most active through the ABTS and FRAP tests, with 184.1 ± 0.67 mg TE/g E and 137.92 ± 0.03 mgET/gE, respectively, while the diethyl ether macerated was the most active via PR test (43.76 ± 0.51 mgEAA/gE). Principal component analysis shows a strong correlation between polyphenols and the FRAP test (r = 0.8369) and flavonoids and the PR test (r = 0.8484). There is a strong correlation between in vitro toxicity, catechic tannin content and reducing power, with r = 0.8079 and r = 0.7869, respectively. Thus, consumption of this mushroom after heat treatment is considered safe and it has an interesting antioxidant potential, which could offer it therapeutic value.

This study investigates the relationship between eight free fatty acids (FFA), triglycerides (TAG), and phosphatidylcholines (PC) after Roux-en-Y gastric bypass (RYGB) to obtain insight into changes in lipid metabolism that might be causally related to postsurgery Type 2 diabetes remission. We measured the FFA, TAG, and PC before and up to 2 years after RYGB in three patient groups: one nondiabetes group (n = 149), one diabetes group with postoperative remission (n = 33), and one diabetes group with persistent diabetes after surgery (n = 20). Pre- and postoperative Pearson's correlations and linear regression models were used to assess the relationship between concentrations of individual FFA, TAG, and PC. At all timepoints, FFA explained more of the variance in PC than in TAG. The level of omega-3 fatty acids was more strongly correlated to TAG in the group with diabetes remission compared to the nondiabetes group. The low plasma concentration of the omega-6 fatty acid dihomo-γ-linolenic acid (DGLA) showed a surprisingly high correlation to both TAG and PC in all three groups. Unexpectedly, in the diabetes group without remission, a nonsignificant relation between saturated stearic acid and monounsaturated oleic acid indicates a possible impaired conversion of stearic acid to oleic acid via the enzyme delta-9-desaturase compared with the two other groups. We conclude that FFA might be involved in regulating the synthesis and metabolism of both TAG and especially PC. Diabetes status might influence the relationship between concentrations of FFA, TAG, and PC, and the data are suggestive of a role for the enzyme delta-9-desaturase in post-RYGB diabetes remission.

Ticks pose a significant global threat to human and animal health as vectors of numerous pathogens, including bacteria, viruses, and parasites. Beyond their harmful impact, tick salivary glands contain serine protease inhibitors (serpins) known for their potential pharmaceutical properties. Traditional methods for studying tick serpins are labor-intensive, but recent advancements in bioinformatics have enabled comprehensive analyses of these molecules. In this study, we employed in silico tools to identify, classify, and analyze serpins encoded within the sialotranscriptome of the camel tick, Hyalomma dromedarii. Through sequence analysis, conserved motifs and domains have been identified, shedding light on evolutionary relationships and functional conservation among serpins both within and between tick species. The complexity of H. dromedarii serpins (HDS) exceeded prior expectations, with the identification of 15 transcripts exclusively expressed in male and 4 transcripts in female H. dromedarii salivary glands, while 91 transcripts are common to both genders. Interestingly, each HDS has a distinct reactive center loop (RCL) sequence at the protein sequence level. These RCL sequences extend from P17 to P4', are similar to those of other serpins, comprise 21 amino acids, and are situated near the C-terminus. All RCLs feature a diverse array of eight amino acid residues at the P1 sites, with the majority (47.06%) having polar basic residues. Moreover, our predictions imply that some HDSs may exert regulatory control over a wide array of proteolytic pathways, indicating their potential involvement in modulating numerous biological processes. Overall, our findings provide valuable insights into tick serpins and lay a solid foundation for future research aiming to exploit these molecules for the development of novel therapeutics and vaccines against tick-borne diseases.

Autoimmune disorders, characterized by an overactive immune system attacking healthy tissues, are a significant global health concern. A common thread among these disorders is vitamin D deficiency. While the exact relationship between vitamin D and autoimmunity remains a hot research arena, plenty of reports have suggested a strong association between vitamin D and autoimmune conditions, including systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, and Type 1 diabetes, among others. Type 1 diabetes, in particular, has been extensively studied in relation to vitamin D. Evidence suggests that vitamin D deficiency may play a role in the development of this autoimmune disease. Beyond its well-known function in bone health, vitamin D acts as a key regulator of both innate and adaptive immune responses. This review sought to explore the complex interplay between vitamin D and the immune system, with a particular focus on its potential role in autoimmunity, especially Type 1 diabetes. Moreover, the coupled therapy and clinical trials that have investigated the use of vitamin D as a therapeutic intervention for autoimmune disorders were also examined, highlighting both successes and limitations.

Gastric cancer (GC) remains a complex disorder with an unclear etiology. Helicobacter pylori, a Gram-negative bacterium, plays a key role in the development of chronic gastritis, peptic ulcers, anemia, and GC. It survives and proliferates in macrophage autophagosomes, gastric epithelial cells, and dendritic cells, with its cytotoxin-associated gene A (CagA) and vacuolating Toxin A (VacA) interfering with autophagy. While most infected patients develop chronic gastritis, 10%-15% develop ulcers, and 1%-3% progress to GC. MicroRNAs (miRNAs), small noncoding RNAs, regulate genes by modulating tumor suppressors and oncogenes, and are crucial in GC. This review explores miRNAs as potential biomarkers and therapeutic targets for early GC diagnosis, monitoring progression, and predicting treatment outcomes. Dysregulated miRNAs such as miR-124, miR-145, and miR-21 have been identified in GC. Additionally, miRNAs influence H. pylori-induced inflammation and the expression of virulence factors. Targeting miRNAs could reduce H. pylori-related diseases, including gastritis, ulcers, and GC. However, further research is required to fully understand miRNAs' role in H. pylori infection and develop miRNA-based therapies.

Plectranthus glandulosus is valued for its culinary and medicinal properties. This study evaluated the cytotoxicity, acute oral toxicity, and subacute oral toxicity of its aqueous extract, hydro-ethanolic extract, and ethyl acetate fraction. Cytotoxicity was tested on African green monkey kidney (Vero) and murine macrophage (RAW 264.7) cell lines exposed to concentrations ≤ 200 μg/mL. Acute toxicity was assessed via a single oral dose of 5000 mg/kg, and subacute toxicity through daily oral doses of 250-1000 mg/kg over 28 days. All extracts maintained > 95% cell viability in Vero and RAW 264.7 cells, indicating no cytotoxicity. No mortality, behavioral changes, or clinical signs of toxicity were observed. Food and water intake, body weight, and organ weights were unaffected (p > 0.05). Macroscopic and histopathological examinations of the liver, kidneys, heart, and spleen showed no abnormalities. At the highest subacute dose (1000 mg/kg), hematological parameters remained within normal ranges, showing no significant differences compared to the control (p > 0.05): white blood cells (7.23-7.95 vs. 7.94 ± 1.29 × 10³/mm³), red blood cells (3.00-3.38 vs. 3.57 ± 0.79 × 10⁶/mm³), hemoglobin (12.10-12.40 vs. 12.20 ± 1.10 g/dL), hematocrit (29.60-44.53 vs. 41.70 ± 3.70%), and platelets (491.30-528.13 vs. 490.70 ± 0.40 × 10³/mm³). Differential leukocyte counts (lymphocytes, monocytes, eosinophils, and basophils) also remained stable. Biochemical markers, including alanine transaminase (62.76-69.20 vs. 64.60 ± 10.25 U/L), aspartate transaminase (112.15-123.85 vs. 120.69 ± 9.56 U/L), total protein (64.45-65.80 vs. 65.2 ± 3.20 g/L), urea (20.35-21.22 vs. 21.62 ± 0.73 mg/dL), creatinine (0.56-0.58 vs. 0.56 ± 0.33 mg/dL), and C-reactive protein (≤ 350 µg/mL), were comparable to controls. In conclusion, P. glandulosus extracts and fraction exhibited no cytotoxic, acute, or subacute toxic effects at the tested doses, supporting their safety and potential in alternative medicine.

Ozone (O₃) and polystyrene nanoparticles (PNPs) display diffusive behavior that leads to toxicity in many tissues of the adult organism. In this study, we evaluated the interactions between atmospheric pollutants and human muscle, using human myogenic progenitor cells (huMPCs) derived from vastus lateralis skeletal muscle. To achieve this goal, O₃ and PNPs were first tested individually to understand the impact of the single pollutant on huMPCs. Subsequently, pollutants were tested in combination to examine their potential synergistic effects, given the simultaneous presence of multiple pollutants in the atmosphere. Cell viability was assessed after treatment with O₃ and PNPs, and it seems to be significantly affected in huMPCs exposed to the pollutants, tested both alone and in combination. Similarly, the differentiation capability of treated huMPCs was evaluated, and it was found to be significantly reduced compared to controls, especially when O₃ and PNPs are tested in combination. Furthermore, an alteration in the expression of microRNAs involved in myogenic cells' proliferation and differentiation pathways was found. In light of the correlation between pollutants and increased oxidant levels, and O₃'s ability to produce the superoxide anion, superoxide anion levels in huMPCs exposed to pollutants were also assessed, and an increase in this oxidant was recorded. Thus, this preliminary study suggests that exposure to O₃ and PNPs affects human muscle, as it alters all the analyzed parameters in huMPCs, filling a gap in the current literature.

Although the World Health Organization has declared that the coronavirus disease (COVID-19) is not a public health emergency of international concern anymore, it has negatively impacted the world, and effective treatment for this pandemic remains a major priority. Vaccine effectiveness has been a matter of concern given the evolution of variants and subvariants of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Thus, continued protection against SARS-CoV-2 and its variants is still necessary and could work alone or in parallel with vaccinations to treat COVID-19 in the future. Further, findings from in vitro and in vivo studies have noted the effectiveness of high dosages of nitric oxide (NO) as an antimicrobial agent against respiratory pathogens such as bacteria, viruses and fungi. NO has been previously utilized in the management of SARS-CoV and has shown a similar antiviral effect with SARS-CoV-2 in vivo and in vitro. Effective therapy with NO can be used to target several stages of COVID-19 infection to prevent transmission and progression of the disease. The unique properties of NO allow this simple, gaseous molecule to be administered in various forms. NO can be used as an inhalant, in the form of NO donor drugs such as S-nitrosothiols and more recently as NO-releasing nanoparticles (NO-nps). This review summarizes the bioavailability of NO in COVID-19 patients and highlights in vivo and in vitro studies as well as clinical trials with NO administered as a nasal spray, inhalant, or via nanodelivery for therapeutic applications for COVID-19 and other respiratory infections in the future.

A heme-containing peroxidase was isolated and characterized from the skin of Cucurbita maxima (PKS). Large-scale purification of the enzyme was performed, yielding a stable and active preparation suitable for detailed biochemical analysis. The PKS's properties were investigated, including pH sensitivity, temperature stability, and the influence of ionic strength on its activity and its spectroscopic properties. Kinetic parameters were determined using fluorophenol substrates and compared to those of the extensively studied horseradish peroxidase (HRP), highlighting similarities and unique features. The primary structure determination of the purified PKS was conducted using protease digestion and subsequent MS fragment analysis. Coupled with genomic data from PKS, a corresponding protein sequence was determined, which was then used to generate a 3-dimensional structure for the PKS enzyme using computational approaches. In addition, a spectrophotometric analysis of the purified PKS was conducted, enabling the determination of molar extinction coefficients for the holoenzyme in its presumed 5-coordinate high-spin ferric state (as isolated) and its 6-coordinate high-spin ferric state upon complexation with fluoride ions. The dissociation constant for fluoride binding exhibited significant pH sensitivity, consistent with observations reported for HRP. Additionally, the PKS enzyme showed distinct spectral binding features when complexed with trifluoroacetic acid, whereas no such binding was observed with HRP. These findings provide insights into the biochemical and structural characteristics of this low-cost plant peroxidase. Furthermore, our results provide relevant information for advancing the potential uses of these peroxidases in biotechnological applications such as environmental remediation of perfluorinated carboxylic acids.