International Journal of Cell Biology最新文献

siRNA is a versatile tool with practical applications in various fields, such as fundamental research, therapeutic development, and plant genetics. A few siRNA therapeutics have been FDA-approved, such as Onpattro (patisiran) and Leqvio (inclisiran) to treat hereditary transthyretin amyloidosis and primary hypercholesterolemia, respectively. In addition, several others are currently in clinical trials, highlighting the potential of siRNA-based treatment for undruggable targets. siRNA is a double-stranded RNA molecule that has the potential to inhibit gene expression by degrading target mRNA. The siRNA sequence must be precisely designed for effective gene knockdown and to minimize off-target effects. Strategies for designing siRNA to achieve optimal efficacy are presented in this review. We emphasize approaches that promote effective gene knockdown by siRNA. These approaches include preventing off-target RNAi and ensuring incorporation of the intended guide/antisense strand into RISC for targeted gene knockdown. This review also discusses the assessment of siRNA efficacy in vitro and the design of appropriate nonsilencing controls. Furthermore, the challenges of in vivo applications are identified, and strategies to overcome these challenges, such as siRNA delivery methods, biodistribution, and immunotoxicity prevention, are highlighted. Lastly, nucleotide chemical modifications to the ribose sugar and phosphodiester bonds and their effects on siRNA stability, activity, and interaction with the RISC complex are discussed. Overall, this review serves as a guide for well-designed and rigorously tested siRNA sequences, starting from initial in silico design to the application of siRNA for research or development of siRNA-based therapeutics.

Background: Lactic acid bacteria (LAB) are extensively used in the preservation of fruits and vegetables due to their ability to inhibit spoilage organisms and pathogenic bacteria. The fermentation process mediated by LAB not only extends the shelf life of produce but also enhances its nutritional and sensory properties. Objective: This study is aimed at reviewing the mechanisms through which LAB preserve fruits and vegetables, evaluating the benefits of their application, and highlighting recent advances in this field. Methods: A comprehensive literature review was conducted, focusing on the various preservative mechanisms of LAB, including acid production, bacteriocin secretion, competition for nutrients, and exopolysaccharide production. The impact of LAB on the shelf life, safety, nutritional value, and sensory attributes of fruits and vegetables was assessed through an analysis of recent scientific studies and case examples. Results: LAB extends the shelf life of fruits and vegetables primarily through the production of lactic acid, which lowers the pH and inhibits the growth of spoilage organisms and pathogens. The secretion of bacteriocins by LAB provides additional antimicrobial activity. Competition for nutrients further suppresses unwanted microbial growth. Exopolysaccharides produced by LAB can improve texture and form protective biofilms. Notable examples of LAB application include the fermentation of cabbage into sauerkraut, the production of kimchi, and the pickling of cucumbers. Advances in biotechnology have led to the development of optimized LAB strains and starter cultures, enhancing the efficacy of preservation processes. Conclusion: LAB are highly effective in preserving fruits and vegetables through multiple mechanisms, offering significant benefits in terms of extended shelf life, improved safety, enhanced nutritional value, and better sensory qualities. Ongoing research and technological advancements are expected to further improve the application of LAB in food preservation, making it a more efficient and widespread practice.

Radiation therapy is one of the most effective treatments for approximately 60% of patients with cancer. During radiation exposure, the overproduction of reactive oxygen species (ROS) disrupts the lipid layer of the membrane, leading to subsequent peroxide radical formation. Cimetidine (Cim) and famotidine (Fam) are histamine H2 receptor antagonists (H2 blocker), also known as peptic ulcer drugs, that exert radioprotective effects. Vitamin C (Vit.C) is an effective free radical and ROS scavenger with significant radioprotective effects. In this experimental study, male mice (6-8 weeks and 28 ± 3 g) were used in five groups. To evaluate ionizing radiation, gamma rays were used at two doses of 2 and 4 Gy and different doses of Cim, Fam, and Vit.C administered as the protectives. Finally, the livers of the mice were isolated and homogenized. The levels of lipid peroxidase and reduced and oxidized glutathione were measured using standard methods. With increasing radiation dose, lipid peroxidase activity, GSSG level, and glutathione content increased. The findings showed that in the drug-only group, Vit.C had better protection than the other two drugs, and the combination of the three drugs had excellent radiation protection. Radiation protection of normal cells in radiotherapy is a valuable necessity. A number of drugs can protect cells against ionizing radiation through different mechanisms. The results suggest that Fam, Cim, and Vit.C can be radioprotective individually or in combination.

Recent research indicates that mesenchymal stem cells (MSCs), known for their anti-inflammatory and anti-infectious properties, could be a promising alternative for treating inflammatory diseases such as inflammatory bowel disease (IBD). This study examined how MSCs and their derivatives, when cocultured with interleukin-1β (IL-1β)-stimulated Caco-2 cells, affect the expression of genes related to inflammation, microbes, and apoptosis. In the experiment, Caco-2 cells were exposed to 10 ng/mL of IL-1β for 24 h. MSCs were sourced from human bone marrow, adipose tissue (AD-MSC), and menstrual blood. These MSCs and their conditioned medium (CM) were then cocultured with the IL-1β-induced Caco-2 cells. After 48 h, gene expression levels were analyzed using real-time PCR, and the data were statistically evaluated using T-tests, U-Mann-Whitney, and Tukey's post hoc analyses. The results indicated that IL-1β at 10 ng/mL was the optimal concentration for inducing Caco-2 cells with the highest viability and minimal damage. Among the MSCs tested, AD-MSCs were the most effective in regulating gene expression. Specifically, AD-MSC treatment significantly reduced the mRNA expression of TNF-α and IL-1β, both of which are crucial in sustaining inflammatory responses (p ≤ 0.05). This study concludes that AD-MSCs have superior effects compared to other MSC sources in modulating genes associated with inflammation, antibacterial effects, and apoptosis in an in vitro model of IBD using Caco-2 cells.

Background: The exponential growth in mobile phone usage has raised concerns about electromagnetic field (EMF) exposure and its health risks. Blood pressure and BMI, which impair heart function due to decreased adrenoreceptor responsiveness, parasympathetic tone withdrawal, and increased sympathetic activity, may further exacerbate these risks. However, the effects of radiofrequency electromagnetic (RF-EM) exposure from mobile phones on electrocardiograms (ECGs) and heart rate variability (HRV) in individuals remain unclear. Purpose: Building upon our previous findings on HRV changes due to mobile phone proximity, this study is aimed at significantly enhancing the analytical approach used to assess the effects of mobile phones on cardiac parameters. This study exploits data from a previous study but with a different purpose. The aim of this study is twofold: (a) to examine whether exposure to mobile phones changes the five variables (P-R, QRS, QT, ST, and HR) in a multivariate manner and (b) to examine whether the blood pressure and/or the body mass index (BMI), which acts as a proxy for obesity, have an effect on the change of these five variables. For both aspects of the study, four cycles are performed. Method: We conducted multivariate analysis on previously collected electrical cardiac measurement data from 20 healthy male subjects exposed to mobile phone EMF, with the mobile phones placed at four different body locations. The one-sample Hotelling T ² test on the mean vector of differences was utilised instead of multiple paired t-tests. This multivariate method comprehensively analyzes data features and accounts for variable correlations, unlike multiple univariate analyses. Given our small sample size, we employed the MMPC variable selection algorithm to identify predictor variables significantly related to mean changes. Results: Significant alterations in ECG intervals and heart rate were noted in the subjects before and after the first EMF exposure cycle, independent of their BMI. Notably, heart rate, P-R, and QRS intervals fell postexposure while QT and ST intervals increased. These changes were influenced by variations in systolic blood pressure, with BMI showing no significant effect. Conclusion: The observed modifications in cardiac electrical measurements due to mobile phone EMF exposure are attributed to the effects of EMF itself, with no impact from BMI on the extent of these changes.

Liver cancer has been reported to be one of the most malignant diseases in the world. It is late diagnosis consequently leads to a difficult treatment, as the cancer reached an advanced stage. Hepatocellular carcinoma (HCC) is the primary type of cancer diagnosed in the liver, with deadly characteristics and a poor prognosis. The first-in-line treatment for advanced HCC is sorafenib. Sorafenib acts by inhibiting cell proliferation and by inducing apoptosis as well as blocks receptors associated with these mechanisms. Due to its constant use, sorafenib resistance has been described, especially to proteins of the Bcl-2 family, and their overexpression of Bcl-XL and Mcl-1. This review focuses on the role of the Bcl-2 proteins in relation to sorafenib resistance as a consequence of first-in-line treatment in HCC.

Elk-1 is a member of the ETS domain transcription factor superfamily that is phosphorylated upon mitogen-activated protein kinase (MAPK) pathway activation, which in turn regulated its interaction with partner protein serum response factor (SRF), leading to formation of a ternary complex with DNA. It has previously been reported that Elk-1 interacts with a mitotic kinase Aurora-A, although the mechanisms or the relevance of this interaction was unclear. Elk-1 was also reported to be phosphorylated by CDK5 on Thr417 residue. In this study, we show for the first time that this transcription factor interacts not only with Aurora-A but also with other mitotic kinases Aurora-B, Plk1, and Cdk1, and we define the interaction domain on Elk-1 to the first N-terminal 205 amino acids. We also describe putative phosphorylation sites of these mitotic kinases on Elk-1 and show that Elk-1 peptides containing these residues get phosphorylated by the mitotic kinases in in vitro kinase assays. We also perform bioinformatic analysis of mitotic phosphoproteomes and determine potential interaction partners for Elk-1 in Plk or Aurora phosphoproteomes. We propose that understanding the dynamic phosphorylation of Elk-1 by mitotic kinases is important and that it can present a novel target for anticancer strategies.

Astrocytes maintain CNS homeostasis but also critically contribute to neurological and psychiatric disorders. Such functional diversity implies an extensive signaling repertoire including extracellular vesicles (EVs) and nanotubes (NTs) that could be involved in protection or damage, as widely shown in various experimental paradigms. However, there is no information associating primary damage to the astrocyte genome, the DNA damage response (DDR), and the EV and NT repertoire. Furthermore, similar studies were not performed on hippocampal astrocytes despite their involvement in memory and learning processes, as well as in the development and maintenance of alcohol addiction. By exposing murine hippocampal astrocytes to 400 mM ethanol (EtOH) and/or 1 μM corticosterone (CTS) for 1 h, we tested whether the induced DNA damage and DDR could elicit significant changes in NTs and surface-attached EVs. Genetic damage and initial DDR were assessed by immunolabeling against the phosphorylated histone variant H2AX (γH2AX), DDR-dependent apoptosis by BAX immunoreactivity, and astrocyte activation by the glial acidic fibrillary protein (GFAP) and phalloidin staining. Surface-attached EVs and NTs were examined via scanning electron microscopy, and labeled proteins were analyzed via confocal microscopy. Relative to controls, astrocytes exposed to EtOH, CTS, or EtOH+CTS showed significant increases in nuclear γlH2AX foci, nuclear and cytoplasmic BAX signals, and EV frequency at the expense of the NT amount, mainly upon EtOH, without detectable signs of morphological reactivity. Furthermore, the largest and most complex EVs originated only in DNA-damaged astrocytes. Obtained results revealed that astrocytes exposed to acute EtOH and/or CTS preserved their typical morphology but presented severe DNA damage, triggered canonical DDR pathways, and early changes in the cell signaling mediated by EVs and NTs. Further deepening of this initial morphological and quantitative analysis is necessary to identify the mechanistic links between genetic damage, DDR, cell-cell communication, and their possible impact on hippocampal neural cells.

Prolonged low-dose administration (PLDA) of several FDA-approved drugs for noncancer conditions or dietary compounds is associated with a lower incidence of specific types of cancers and with the lower formation of metastasis. However, the underlying mechanism is unknown; there is a discrepancy between the concentration of drugs needed to kill cancer cells in vitro and the actual serum levels (10 and >1000 times lower) found in patients. In this study, we evaluated the hypothesis that clonogenicity may be the target of PLDA. We compared the effect of nigericin (NIG) and menadione (MEN) on the human A549 and H460 lung and MCF-7 and MDA-MB-231 breast cancer cell lines using routine MTT and colony forming assays (CFA). The ability of both NIG and MEN to eliminate 100% of cancer cells was at least 2-10 times more potent in CFA compared to MTT assays. Our results revealed the existence of a short post-reattachment window of time when cancer cells growing at low density are more sensitive to PLDA of specific drugs likely by targeting clonogenic rather than proliferation pathways. This short ultrasensitive window of time (SUSWoT) was cell- and drug-type specific: the SUSWoT for NIG was present in H460, A549, and MDA-MB-231 cells but not evident in MCF-7 cells. Conversely, a similar SUSWoT for MEN was present in MCF-7, MDA-MD-231, and A549 cells but not evident in H460 cells. Our findings partially explain the decreased incidence of specific types of cancer by PLDA of FDA-approved drugs (or dietary compounds) for noncancer conditions.

Inducible gene regulation methods are indispensable in diverse biological applications, yet many of them have severe limitations in their applicability. These include inducer toxicity, a limited variety of organisms the given system can be used in, and side effects of the induction method. In this study, a novel inducible system, the RuX system, was created using a mutant ligand-binding domain of the glucocorticoid receptor (CS1/CD), used together with various genetic elements such as the Gal4 DNA-binding domain or Cre recombinase. The RuX system is shown to be capable of over 1000-fold inducibility, has flexible applications, and is offered for use in cell cultures.