Cellular and molecular biology最新文献

This study explores a novel approach to treating endodontic infections by targeting the acetate kinase (Ack) enzyme of Porphyromonas gingivalis, a key pathogen in these infections. Using computational methods, we developed an apo receptor-based E-pharmacophore model of P. gingivalis Ack and screened the ZINC Lead-Like database containing over 1.8 million compounds. High-throughput virtual screening and molecular dynamics simulations identified ZINC001306857494 as a promising lead compound, demonstrating stable binding to the Ack active site with a binding free energy of -41.66 kcal/mol. The compound forms multiple hydrogen bonds with highly conserved residues, including Leu119, His180, and Arg241. Molecular dynamics simulations over 250 ns confirmed the stability of the protein-ligand complex, with sustained interactions throughout the simulation period. This study presents a potential new scaffold for developing Ack inhibitors, offering a promising avenue for treating endodontic infections caused by P. gingivalis.

The present study explores the simple and eco-friendly green synthesis of silver (AgNPs) and gold (AuNPs) nanoparticles using aqueous flower extract of Bombax ceiba, commonly known as silk cotton. The extract, rich in flavonoids, serves as both a reducing and capping agent, facilitating the synthesis of metal nanoparticles. The synthesized nanoparticles were characterized using UV spectroscopy and scanning electron microscopy (SEM), confirming their formation and stability. The antibacterial activity of the AgNPs, AuNPs, and crude flavonoids was evaluated against several bacterial strains, including Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus, using the agar well diffusion method. Our results show that AgNPs exhibit significant antibacterial activity, particularly against Gram-negative bacteria, with a marked zone of inhibition observed for S. typhi and E. coli. The inhibition zone increased with higher concentrations of AgNPs. In contrast, AuNPs and flavonoid solutions demonstrated only mild antibacterial effects, with no significant inhibition observed at lower concentrations (1-6 µL). The antibacterial efficacy of AgNPs was comparable to that of standard antibiotics, such as Azithromycin for Gram-positive bacteria and Ciprofloxacin for Gram-negative bacteria, suggesting their potential as effective antimicrobial agents. The antibacterial activity of the synthesized nanoparticles and the crude flavonoid extract highlights the promising use of Bombax ceiba flower extract in the green synthesis of metal nanoparticles with potential biomedical applications.

The development of treatment methods used in the treatment of non-small cell lung cancer (NSCLC) is important to prevent problem of increasing mortality. However, the treatment methods used in clinical settings at the clinic are insufficient to eliminate this problem. For this purpose, it was aimed to determine whether the combination of docetaxel (DTX) and FH535 can be used as an anticancer agent candidate in A549 cells and whether it is a candidate drug combination that can be used in clinical treatment after in vivo studies. FH535 is a WNT signaling pathway inhibitor and is known to be overactive in NSCLC. In this study, the effects of DTX and WNT signaling pathway inhibitor FH535 used in NSCLC treatment on A549 and BEAS-2B cell lines were evaluated at the cellular level. While increasing the anticancer activity in A549 cells, the doses showing minimum toxic effect in BEAS-2B cells were determined by Real Time Cell Analysis method. Mitotic activity, BrdU cell proliferation assay and caspase 3,7 activity assay were performed for detailed analysis of the combination dose at cellular level. The results show that the combined dose had an antimitotic effect on A549 cells, causing mitotic catastrophism, while in BEAS-2B cells neither agent was more toxic than either agent alone, reducing mitotic activity and BrdU activity, leading the cell to mitotic catastrophism, while caspase 3,7 activity was unchanged. This study demonstrated for the first time the effects of the combination of DTX and FH535 on A549 and BEAS-2B cell lines.

Nephrotoxicity is characterized by the adverse effects on kidney function caused by various substances, including hazardous chemicals and drugs. This study aimed to investigate the neuroprotective properties of aqueous, methanolic, and ethanolic extracts of Tamarix dioica leaf against acetaminophen-induced kidney damage and compare their efficacy with metformin, a known neuroprotective agent. Thirty-six albino mice were randomly divided into six groups, including a standard control group, an acetaminophen-toxified group, a positive control group treated with metformin (at a dose of 200 mg/kg body weight), and three experimental groups treated with aqueous, methanolic, and ethanolic T. dioica extracts (at a dose of 400 mg/kg body weight each). The neuroprotective potential of the T. dioica extracts was assessed by evaluating hematological markers, electrolyte levels (Na+, K+, Cl-), antioxidant enzymes (CAT, SOD, MDA), renal function tests (urea and creatinine), and toxicity markers (SGOT and SGPT). Additionally, histopathological analysis was conducted to observe any pathological changes in kidney tissues stained with hematoxylin and eosin. The results demonstrated that the T. dioica leaf extracts effectively restored all the indicators and antioxidant enzyme levels to normal, significantly differing from the elevated levels observed in the acetaminophen control group (p < 0.05). Furthermore, histopathological examination revealed regeneration of glomeruli and renal tubules in the stained tissues. These findings suggest that T. dioica leaf extracts can potentially mitigate acetaminophen-induced nephrotoxicity.

Chondroitin Sulfate (CS) is an essential component of the extracellular matrix and is a sulfated glycosaminoglycan structurally composed of a polysaccharide chain consisting of N-acetyl galactosamine and glucuronic acid. The use of CS of animal origin is common in pharmacological research. The disadvantages of traditional sources and methods used in the production of CS, which is used in various applications in the medicine, veterinary, pharmacy, and cosmetic sectors, have made microbial production a vital alternative. In this study, recombinant Escherichia coli (pETM6-PACF-vgb) strain, in which kfoA, kfoC, kfoF and vgb gene regions are co-expressed, and E. coli pETM6-PACF strain, which does not contain the vgb gene, were used in the microbial production of CS. The vgb gene is the region responsible for expressing the bacterial protein Vitreoscilla hemoglobin (VtHb). This study investigated the effect of the expression of VtHb in E. coli on increasing bacterial cell respiration and, therefore, how ATP production would affect cell growth and the acquisition of chondroitin and microbial chondroitin sulfate (MCS) from biomass. The analysis results determined a 23.07% increase in the amount of MCS produced from the vgb+ strain. The presence of vgb had positively affected culture age and reproductive kinetics. Spectrophotometric measurements, NMR, HPLC, FT-IR, TGA, DTA, and DSC analyses for the reproductive values ​​and physicochemical characterization of the obtained MCS were applied to discuss this production process. For more detailed results on this subject, future research focused on optimization is needed.

Current research reveals a complex relationship between blood cells(BC) and neuropathic pain(NP), though the underlying biological mechanisms remain unclear. This study applies Mendelian randomization (MR) to investigate causal relationships between BC and three major types of NP: diabetic peripheral neuropathy(PDPN), postherpetic neuralgia(PHN), and trigeminal neuralgia(TN). We also explore the potential mediating role of immune cells in these associations. We employed a two-sample, two-step Mendelian randomization study using the inverse variance weighted method to investigate the causal effect of BC on three major types of NP, as well as the mediating role of immune cells in the association between BC and NP. Additionally, we utilized a two-step Mendelian randomization design to explore the mediating effect of immune cells. We identified 13 distinct blood cell phenotypes under various perturbation conditions that have a significant causal relationship with NP. Additionally, we discovered 127 immune cells that exhibit a notable causal connection with NP. Through Mendelian Randomization (MR) and two-step Mendelian Randomization analyses, we found the following results: Three blood cell phenotypes were associated with PDPN, three with PHN, and seven with TN, with platelet, red blood cell, monocyte, and neutrophil responses showing significant correlations with NP risks. Immune cell analyses revealed 36 phenotypes increasing and 31 decreasing PDPN risk, 16 increasing and 21 decreasing PHN risk, and 18 increasing and 13 decreasing TN risk, with HLA DR on DCs, PB/PC AC, and CD39+ CD4+ %T cell showing the strongest associations, respectively. Mediation analysis identified immune cells, such as CD39+ resting Treg and HLA DR+ CD4+ %lymphocyte, mediating PBC effects on NP risks. Sensitivity analyses confirmed no significant heterogeneity or pleiotropy, and reverse MR analyses found no reverse causal relationships. This study provides new evidence for the causal relationship between blood cell phenotypes and neuropathic pain and proposes immune factors with potential mediating effects. However, this finding needs to be further demonstrated by more extensive clinical studies.

Interleukin-33(IL33), an alarm cytokine of the IL-1 family, is expressed mainly in epithelial cells of barrier tissues and is involved in the repair of epithelia to maintain barrier function. However, the mechanisms regulating IL33 expression and the mechanisms by which IL33 regulates the intestinal barrier function are not fully clarified. In this study, Caco-2 cells and siRNA were applied to investigate the role of Notch/IL33/ST2 Signaling in regulating intestinal epithelial barrier function, which was demonstrated by protein expression of tight junctions and trans-epithelial resistance (TER) assay. Our results revealed that Notch signaling pathway was activated and IL33 expression was up-regulated after LPS stimulation. After blocking Notch signaling with DPAT or siRNA for Notch1, IL33 expression was significantly down-regulated in Caco-2 cells. The protein expression of tight junctions (ZO-1, occludin, and claudin-1) was down-regulated after siRNA for IL33 in Caco-2 cells with LPS stimulation. Also, the intestinal epithelial TER was down-regulated after siRNA for IL33 with LPS stimulation or not. Exogeneous IL33 promoted the tight junction protein expression and increased the TER. Finally, our data further showed that IL33 regulates intestinal epithelial barrier function through the ST2 receptor. In conclusion, our results indicated that IL33/ST2 axis, which was activated by the Notch signaling, maintains intestinal epithelial barrier function through regulating tight junction protein expression under inflammatory conditions. This study provides a new therapeutic pathway of regulating intestinal epithelial barrier dysfunction.

Widespread human exposure to phthalates is caused by their intensive usage in industrial and consumer plastic products. DEHP (di(2-ethylhexyl) phthalate) is one of the most often used phthalates and is presented not only in food and fluids but also in the air and dust contact with plastic products. Regrettably, phthalates easily migrate into the human body and act as potent toxicants, mainly on endocrine and metabolic status. In the last decade, several epidemiological studies have indicated a correlation between prenatal exposure to phthalates and adverse effects on neurodevelopment in offspring. Our research aimed to assess the impact of DEHP prenatal subchronic exposure on male offspring's behavior and learning ability and identify the primary target brain structure/s of neurotoxic action. Heightened anxiety in male offspring was evident through increased rearing, frequent line crossings, hurried movements, and reduced grooming behavior. These behaviors were accompanied by a decline in recognition memory and diminished interest in exploring novel objects. Obtained data showed that prenatal oral exposure to DEHP in a selected concentration induces irreversible changes in brain structures of the male offspring, primarily in the hippocampus, that underlies significant alterations in cognitive behavior and enhanced anxiety. The molecular mechanism of DEHP-induced hippocampal neurotoxicity in the maturing male brain involves changes in phosphatase and tensin homolog (PTEN) subcellular location, which suppresses Akt/mTOR signaling, enhances GluN2B NMDA mediated synapse depression and decreases mitochondrial fusion.

The liver is the second largest organ in the body, playing a crucial role in maintaining homeostasis and regulating metabolism. However, the prevalence of liver diseases has been rising steadily due to an increase in both infectious and non-infectious factors. Medicinal plants offer a remarkable opportunity to enhance our healthcare system by addressing various diseases through their ability to control oxidative stress and support metabolic processes. This revision improves readability while retaining the original meaning. In the present study, purified methanol extracts of Martynia annua and Polygonum viviparum at different concentrations were used to explore the antioxidant and hepatoprotective potential against abiotic stress on liver cells.  Firstly total flavonoids and total phenolic contents of both plants were measured and then their antioxidant activities were determined through DPPH radical scavenging activity and FRAP assay. Moreover, bioactive compounds and in vitro hepatoprotective potential among these plants were determined through LC-MS and Liver Slice Culture assay respectively. In P. viviparum high amounts of TPC and TFC were observed with maximum antioxidant potential in terms of DPPH inhibition at 84% and high ferric-reducing ability at 240 mg/mL. The presence of different phytoconstituents like Myricetin, Gallic acid, Ferulic acid, Chlorogenic acid, Sweroside, Morroniside, Echonoside, Swertiamarin and Protocatechuic acid were confirmed in M. annua and P. viviparum in LC-MS study. The maximum hepatoprotective potential in terms of minimum cytotoxicity 6% and 9% were observed in M. annua and P. viviparum respectively. It was revealed that both plants have stunning hepatoprotective properties to prevent liver from toxicants and their related complications due to high antioxidant potential and active metabolites.

Pompe disease which is glycogen storage disease type II, is an autosomal recessive lysosomal storage disorder where GAA gene mutations cause deficiency of acid alpha-glucosidase leading to deposition of glycogen in various tissues. Chromosome 17q25.2-25.3 is the location of GAA gene. This study aims to collect information on the Pompe disease symptoms' severity and genotypes of 18 children who represented all infant patients with Pompe disease until March 1st, 2024. For diagnosis tandem mass spectrometry and genetic study were used. Muscle strength was assessed by hand-held dynamometry. Cardiac assessment was by echocardiography and electrocardiography. The feeding and swallowing difficulties in the patients were addressed. Statistical analysis was used P<0.05 was considered significant. Fifty percent had normal mental development, 27.8% had delayed mile stones 55.6% had weakness of extremities, 50% had heart problems in the first month,38.8% had respiratory problems in the first month and 12(66.6%) had feeding difficulties. The level of the enzyme alpha-1,4 Glucosidase level was Zero in two patients 66.7% and was 0.1µmol/L/h in 33.3% of the alive patients while it was 0.1 µmol/L/h in 73.3% and 0.2 µmol/L/h in 13.3% of the dead with a significant correlation. The genetic mutations were   c. [258dupC]; [258dup] in 6 (33.3%) of the patients, c.258dup in 3(16.6%) and c.2237G>A in 11.1% of all the patients. Childhood Pompe disease course varies widely. It is important to consider Pompe disease in the differential diagnosis of patients with unexplained fatigue and weakness and cardiorespiratory involvement.