Reports of Biochemistry and Molecular Biology最新文献

Background: 3,4-methylenedioxymethamphetamine (MDMA) affects the male reproductive system. We investigated the mRNA levels of bax, bcl-2, and p53 genes in MDMA-induced apoptosis in mouse Sertoli (TM4) and Leydig (TM3) cells before and after taking Zinc.

Methods: The TM3 and TM4 cells were cultured in four groups: I (untreated medium), II (medium with 5 mM MDMA), III (medium with 8 µM Zinc), and IV (medium with 8 µM Zinc prior to 5 mM MDMA administration). After 48 hours, total RNA was extracted from the samples, and cDNA was synthesized. The relative gene expression level was evaluated using the SYBR Green PCR kit.

Results: In the MDMA group, the relative amounts of bax and p53 gene expressions increased; conversely, the relative amount of bcl-2 gene expression decreased in TM3 and TM4 cell lines. In the MDMA+Zinc group, there were no statistically significant differences between this group and the control group regarding the mRNA levels of bax and p53 genes in the TM3 cell line, as well as the mRNA levels of bax and bcl-2 genes in the TM4 cell line. Statistically significant differences were observed between the MDMA+Zinc and MDMA groups regarding the relative expressions of bax and p53 genes in the TM3 cell line and bcl-2 and p53 genes in the TM4 cell line.

Conclusions: Zinc mitigates MDMA- induced apoptosis by altering the crosstalk among bcl-2, bax, and p53 gene expressions in the tested cell lines.

Background: The oral delivery of therapeutic peptides and proteins presents a significant challenge in pharmaceutical development due to barriers such as the intestinal epithelium and the blood-brain barrier (BBB). These barriers limit the passage of large, hydrophilic molecules through transcellular pathways and restrict paracellular transport due to intercellular tight junctions. This study investigates the potential of E- cadherin-modulating peptide, ADT-6, to improve the penetration of these therapeutic agents.

Methods: We constructed a fusion protein of ADT-6 and green fluorescent protein (GFP) to evaluate its activity and transport through the epithelial cells' paracellular pathway. Using Escherichia coli strains for expression, we cloned the GFP-ADT-6 construct, which provides a solid foundation for our study's methodology.

Results: Our molecular simulations showed that the linker between GFP and ADT-6 maintains the fusion protein's integrity and provides flexibility in receptor interaction. Permeability experiments revealed that ADT-6 markedly reduced transepithelial electrical resistance (TEER) and significantly increased GFP transfection in Caco-2 cell monolayers dose-dependently. Results of ELISA confirmed these findings, showing high GFP levels in the lower compartment of Transwell systems treated with GFP-ADT-6.

Conclusions: This study demonstrates the potential of ADT-6 to deliver proteins from the paracellular route, enhance the bioavailability of pharmaceutical drugs by altering cell-cell interactions, and provide new opportunities for oral drug delivery strategies.

Background: Systemic inflammation is one of hallmarks in chronic obstructive pulmonary disease (COPD), contributing to high morbidity and mortality due to elevated levels of interleukin-6 (IL-6) and reduced level of interleukin-10 (IL-10). Probiotics have the potential to reduce systemic inflammation through the gut-lung axis. This study aims to assess the effect of probiotics compared with an inhaled bronchodilator on serum IL-6 and IL-10 levels in mice model of COPD.

Methods: This was an in vivo experimental study with a post-test only control group design. Thirty C57BL/6 mice were randomized into five groups; NC (healthy mice), PC (COPD induced mice); T1 (COPD mice treated with a bronchodilator), T2 (COPD mice treated with probiotics) and T3 (COPD mice treated with both a bronchodilator and probiotics). COPD was induced for 12 weeks, followed by a 6-week treatment period. After completing the treatment, serum IL-6 and IL-10 levels were measured using the enzyme-linked immunosorbent assay (ELISA).

Results: The IL-6 levels in T2 group were reduced to levels comparable to the negative control group (13.5 vs 12.0 pg/ml respectively, p=0.84). The IL-10 levels were higher in T2 group compared to T1 group, however; this difference was not statistically significant (181.4 vs 155.0 respectively, p>0.05).

Conclusions: In mice model of COPD, probiotics have been shown to lower IL-6 levels and, to a lesser extent, increased IL-10. As a result, probiotics may have a protective effect against systemic inflammation.

Background: One of the deadliest cancers in the world, colorectal cancer has a dismal prognosis and a poor response to therapy. It was suggested that outer membrane vesicles (OMVs) produced by Escherichia coli (E. coli) are a powerful inducer of inflammation in intestinal epithelial cells. This research aimed to determine the anticancer potential of E. coli-derived OMVs using a colorectal cancer model.

Methods: Five distinct E. coli strains were collected for this study. Their OMVs were then isolated and characterized using dynamic light scattering (DLS) and scanning electron microscopy (SEM). The effects of E. coli-derived OMVs on colorectal cancer were evaluated in vitro and in vivo using a colorectal tumor model in nude mice.

Results: Obtained results showed that E. coli probiotic strains released spherical-shaped vesicles ranging from 5 to 200 nm. E. coli-derived OMVs showed that in the untreated group, a large portion of the tumor tissue continued to grow, with only a few cells undergoing apoptosis. Conversely, the OMV-treated group exhibited a higher number of apoptotic cells, highlighting the anticancer effects of E. coli-derived OMVs in colorectal cancer.

Conclusions: These results demonstrated that E. coli-derived OMVs can be employed as a potential treatment for colorectal cancer with minimal adverse effects. Mechanistic studies indicate that these vesicles may promote apoptosis and inhibit cell proliferation, supporting their therapeutic potential.

Background: Medicinal plants play an important role in agricultural production due to their therapeutic significance, particularly in the treatment of various pathological conditions.

Methods: Active compounds in Eruca sativa were identified using High-Performance Liquid Chromatography (HPLC). Fifty isolates of Staphylococcus aureus (S. aureus) bacteria were obtained. The effect of Eruca sativa plant extract on biofilm formation of bacterial isolates was tested using the standard plate method.

Results: The highest percentage of S. aureus was found in wound samples, with 22 isolates (44%). The isolates showed variability in their ability to form biofilms. The efficacy test revealed that plants treated with different concentrations of brassinolide (0, 1.5, 2.5, 3.5 mg/L) and Eruca sativa showed inhibition of S. aureus growth in isolates (S2, S7, S10, S15, S16, S31, S42, S48, S50) with alcohol concentrations (5, 25, 75 mg/ml). The effect of Eruca sativa plant extract was dependent on the concentration of brassinolide applied. The highest inhibition was observed with brassinolide concentration of 3.5 mg/L-1 and alcohol extract concentrations of 50 and 75 mg/ml.

Conclusions: The alcoholic extract from the leaves of Eruca sativa, combined with brassinolide, proved effective in inhibiting the growth and biofilm formation of S. aureus bacteria.

Background: This study aimed to synthesize copper oxide (CuO) and silver oxide (Ag2O) nanoparticles using a green synthesis method involving moringa extract and incorporate them into polymer nanocomposites with polyacrolein. The objective was to evaluate their cytotoxicity against fibroblasts and glioblastoma cell lines.

Methods: The CuO and Ag₂O nanoparticles were synthesized using moringa extract as a reducing agent. Nanocomposites were formed through a condensation reaction with polyacrolein. Characterization techniques included Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FT-IR), Transmission Electron Microscopy (TEM), X-Ray Diffraction (XRD), Thermogravimetric Analysis (TGA), and Differential Scanning Calorimetry (DSC). Cytotoxicity was evaluated through in vitro assays using human dermal fibroblasts (HdFn) and A172 glioblastoma cells.

Results: AFM analysis showed nanoparticle sizes of 19.36 nm for Ag2O and 66.89 nm for CuO, while TEM images revealed nonhomogeneous spherical nanocomposites. FT-IR and XRD confirmed the successful incorporation of nanoparticles into the polymer matrix. TGA and DSC results demonstrated thermal stability and transitions of the nanocomposites. Cytotoxicity assays indicated significant inhibition of A172 glioblastoma cell proliferation with minimal impact on normal fibroblast cells, suggesting selective cytotoxicity.

Conclusions: The polymer nanocomposites incorporating moringa-extracted CuO and Ag2O nanoparticles exhibited promising selective cytotoxicity against glioblastoma cells, indicating their potential use as anticancer agents. Further studies on in vivo applications and long-term stability are warranted to advance their biomedical use.

Background: Congenital heart defects (CHD) are recognized as the most common heart abnormalities amongst newborns and children, and atrial septal defect (ASD) is recognized as one of the most frequent forms of CHD. Prior studies indicated that the methylenetetrahydrofolate reductase (MTHFR) gene contributes to the etiology of CHD. Therefore, we designed a case-control study to assess the possible role of the MTHFR gene, specifically the C677T (rs1801133) and A1298C (rs1801131) polymorphisms within the Iranian ASD population sample.

Methods: A total of 166 subjects (81 children diagnosed with ASD and 85 control participants) were enrolled in this research. Samples genotyped for MTHFR rs1801133 and rs1801131 polymorphisms using the PCR-RFLP and ARMS-PCR approaches.

Results: Our results indicated that rs1801131 variant reduced the risk of ASD in codominant (OR [95%CI]: 0.41[0.21-0.83], P=0.012), dominant (OR[95%CI]: 0.48 [0.25-0.93], p=0.028) and overdominant (OR[95%CI]: 0.44 [0.23-0.81], P=0.009) models. Moreover, rs1801133 variant increased the risk of ASD in codominant (OR[95%CI]: 2.68[1.39-5.16], P = 0.003), dominant (OR [95% CI]: 2.72 [1.43-5.14], P = 0.002), overdominant (OR [95% CI]: 2.50 [1.31-4.78], P = 0.005), and allelic (OR [95% CI]: 2.16 [1.27-3.69], P = 0.004) models.

Conclusions: Our findings suggest that MTHFR rs1801133 and rs1801131 variants may potentially affect the onset of ASD.

Background: Type 2 diabetes mellitus (T2DM) poses a significant public health challenge due to its high prevalence. Diabetic nephropathy (DN) is one of the most severe complications associated with T2DM. Early prediction of DN in patients with T2DM can significantly aid in managing this disease. This study takes an approach by investigating the potential role of melatonin and thyroid hormone levels as predictive biomarkers for the progression of diabetic nephropathy in individuals diagnosed with type 2 diabetes mellitus.

Methods: Our cross-sectional study involved 120 male participants, divided into two groups: 60 patients with T2DM and 60 with DN. The Cobas technique was used to measure serum thyroid hormone levels and quantified melatonin levels using an enzyme-linked immunosorbent assay (ELISA). A receiver utilizing characteristic (ROC) curve analysis to evaluate the predictive value of serum melatonin for DN was performed.

Results: No notable disparities in thyroid function tests were observed between diabetic patients with and without DN. However, the average serum melatonin quantity in patients with DN. (177.25 ± 60.48 pg/mL) was drastically lower in those with T2DM without DN (199.9 ± 55.16 pg/mL). The sensitivity and specificity of melatonin in predicting DN were 78% and 76%, respectively, with an optimal cut-off value of 178 pg/mL.

Conclusions: Serum melatonin levels exhibited a notable reduction. among individuals who were diabetic with DN, suggesting its potential utility as an additional predictive marker for developing DN in patients with T2DM.

Background: Type 1 diabetes mellitus (T1DM) is a chronic autoimmune condition that can lead to long-term complications due to oxidative stress and metabolic dysregulation. Paraoxonase-1 (PON-1), an enzyme associated with high-density lipoprotein (HDL), has dual activities: arylesterase and lactonase. These activities protect lipids from oxidative damage. The functional status of PON-1 in children with T1DM may provide insights into the relationship between oxidative stress and the enzyme's protective role. This study aims to assess the arylesterase and lactonase activities of PON-1 in Iraqi children with T1DM.

Methods: Sixty-seven children with T1DM were enrolled and compared with 57 age-matched healthy controls. The enzymatic activities of arylesterase and lactonase were measured to evaluate PON-1's functional status. The Paraoxonase-1/HDL (PON/HDL) ratio was calculated to assess lipid protection and antioxidant capacity. Oxidative status was assessed by measuring total oxidative status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI).

Results: PON-1 activity analysis showed a significant reduction in arylesterase (2.36 ± 1.17) and lactonase (21.9 ± 7.31) in the patients group compared to controls (arylesterase=4.54 ± 1.84, lactonase =29.51 ± 9.92). TOS and OSI were significantly higher, while TAS was significantly lower in the patients group. Pearson correlation revealed a positive correlation between HDL-C and arylesterase (P = 0.002, r = 0.379), and HDL-C and lactonase (P = 0.040, r = 0.366).

Conclusions: Reduced PON-1 activity is associated with T1DM, suggesting that enhancing PON-1 or reducing oxidative stress may help prevent diabetic complications and improve cardiovascular health.

Background: Fusobacterium nucleatum (F. nucleatum) is known to increase in number under hyperglycemic conditions, as it is thought to utilize glucose as a nutrient source. The process of glucose utilization in bacteria occurs with the assistance of enzymatic proteins such as glucokinase. This study aims to investigate the glucose utilization by F. nucleatum ATCC 25586 by examining its growth in glucose-enriched media and its relationship with protein activity through FTIR analysis.

Methods: F. nucleatum ATCC 25586 was cultured in media enriched with 2%, 1%, 0.75%, 0.5%, and 0.25% glucose. Its growth was measured using a spectrophotometer, and protein activity was assessed with FTIR at 24 and 48 hours of incubation.

Results: The results showed that F. nucleatum could utilize glucose as a nutrient source, indicated by growth and protein activity. The maximum growth of F. nucleatum occurred at a 0.75% glucose concentration at 24 hours. However, the Kruskal-Wallis test showed no significant differences in the growth and protein activity of F. nucleatum across the five glucose concentrations (growth, p =0.271 and protein, p =0.149). Spearman correlation analysis indicated no correlation between the growth and protein activity of F. nucleatum (p=0.323). The protein activity of F. nucleatum remained stable across various growth levels.

Conclusions: It can be concluded that glucose could influence the growth of F. nucleatum, although the growth and protein activity of the bacteria did not differ significantly based on glucose concentration. F. nucleatum grown in various glucose concentrations exhibits stable protein activity.