Search Results (258)

Background: During infection with the cytomegalovirus (CMV), the membrane system of the infected cell is remodelled into a megastructure called the assembly compartment (AC). These extensive changes may involve the manipulation of the host cell proteome by targeting a pleiotropic function of the cell such as ubiquitination (Ub). In this study, we investigate whether the Ub system is required for the establishment and maintenance of the AC in murine CMV (MCMV)-infected cells Methods: NIH3T3 cells were infected with wild-type and recombinant MCMVs and the Ub system was inhibited with PYR-41. The expression of viral and host cell proteins was analyzed by Western blot. AC formation was monitored by immunofluorescence with confocal imaging and long-term live imaging as the dislocation of the Golgi and expansion of Rab10-positive tubular membranes (Rab10 TMs). A cell line with inducible expression of hemagglutinin (HA)-Ub was constructed to monitor ubiquitination. siRNA was used to deplete host cell factors. Infectious virion production was monitored using the plaque assay. Results: The Ub system is required for the establishment of the infection, progression of the replication cycle, viral gene expression and production of infectious virions. The Ub system also regulates the establishment and maintenance of the AC, including the expansion of Rab10 TMs. Increased ubiquitination of WASHC1, which is recruited to the machinery that drives the growth of Rab10 TMs, is consistent with Ub-dependent rheostatic control of membrane tubulation and the continued expansion of Rab10 TMs. Conclusions: The Ub system is intensively utilized at all stages of the MCMV replication cycle, including the reorganization of the membrane system into the AC. Disruption of rheostatic control of the membrane tubulation by ubiquitination and expansion of Rab10 TREs within the AC may contribute to the development of a sufficient amount of tubular membranes for virion envelopment. Full article

The present investigation was conducted to observe the effects of different energy densities of a low-level red laser (LLRL) on human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs). hESC-MSCs were cultured and irradiated with a LLRL from 0.5 to 5.0 J/cm² at a wavelength of 635 nm. Biological parameters such as proliferation, viability, and migration were observed after 72 h of LLRL irradiation. Compared with the control, LLRL irradiation significantly increased the proliferation and viability of hESC-MSCs from 0.5 to 2.5 J/cm² (p < 0.001, p < 0.05). LLRL irradiation from 0.5 to 3.0 J/cm² significantly increased the migration of hESC-MSCs (p < 0.01). These results revealed that LLRL irradiation at lower energy densities significantly increased the proliferation, viability, and migration of hESC-MSCs. However, higher energy densities were ineffective; this was also true when we examined osteogenic differentiation, as low energy densities of LLRL had a positive effect on differentiation, whereas higher energy densities had a negative effect on alkaline phosphatase activity, Alizarin Red staining and gene expression analysis. In addition, not all stem cell markers were affected by the laser, and a slight decrease in the expression of CD146, which is a stemness marker, was detected, indicating improved differentiation. These findings indicate that low energy densities of LLRL irradiation have positive effects on the proliferation, migration, and differentiation of hESC-MSCs. However, higher energy densities showed inhibitory effects. Full article

Intra-tissue endoscopy, providing real-time images at all scales, from macroscopic to microscopic, from inside living tissue during surgical procedures, has revealed the existence of a body-wide fibrillar architecture that extends from the surface of the skin to the cell. Different types of cells are housed within this fibrillar architecture and gather together to carry out specific functions. This challenges the commonly accepted notion of the organization of living matter that associates separate organs with connective tissue packaging. We are thus confronted with the global nature of the living human body and its vital processes. This paper sets out to describe the architecture of this fibrillar network which could be assimilated with the fascial tissue and which attributes a more constitutive role to connective tissue. It also demonstrates how movements within this fibrillar network can occur with minimal local distortion while maintaining tissue continuity. The authors propose that the gliding of tissues can be explained by the existence of a highly adaptable fibrillar network that enables the gliding of distinct anatomical structures such as tendons and muscles, without any dynamic influence on the surrounding tissues. The authors propose a new model of tissue movement based on the observation of a ubiquitous dynamic polyhedric fibrillar network with an apparently dispersed and complex pattern of organization, that forms fluid-filled microvolumes, and is found everywhere in the human body. Furthermore, this fibrillar network appears to act as a force absorption system, in addition to providing a framework or scaffolding for cells throughout the body. Observation during intra-tissue endoscopy suggests that this fundamental architectural organization extends into the extracellular matrix that is the natural environment of all cells in the living body, regardless of their size, location or specific function. Full article

Introduction: Lipid metabolism plays a crucial role in breast cancer’s progression, treatment response, and prognosis. Alterations in triglycerides (TGs), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) have been implicated in tumor aggressiveness and chemotherapy outcomes. This review examines the relationship between dyslipidemia and breast cancer, with a focus on chemotherapy-induced lipid alterations and their prognostic significance. Methods: A comprehensive literature search was conducted in PUBMED, Web of Science, and Google Scholar, identifying 108 unique studies. After applying the inclusion criteria, 21 studies were selected for analysis, covering lipid profile changes before, during, and after chemotherapy, as well as their impact on treatment response and clinical outcomes. Results: Breast cancer patients exhibited lower baseline TC, TG, and LDL-C levels compared to healthy controls; however, chemotherapy significantly increased these markers while decreasing HDL-C from 1.1 to 0.9 mmol/L. The incidence of dyslipidemia rose from 42.98% pre-treatment to 58.28% post-treatment. Chemotherapy-induced lipid alterations were most pronounced in anthracycline- and taxane-based regimens, leading to a 38% increase in TGs and a 23% reduction in HDL-C. While some studies reported that lipid levels normalized post-treatment, others indicated persistent dyslipidemia up to 12 months later. High baseline HDL-C was associated with a better chemotherapy response, whereas elevated TGs and LDL-C correlated with increased tumor aggressiveness, lower pathological complete response rates, and a higher relapse risk. Patients with persistently high post-treatment TGs had significantly worse disease-free survival, with a 30% relapse rate compared to 18% in those with normal TG. Preliminary evidence suggests that lipid-lowering therapies, such as statins, may offer therapeutic benefits in breast cancer by targeting the cholesterol synthesis pathways involved in tumor growth, though further clinical trials are required. Conclusions: Dyslipidemia is a key metabolic factor influencing breast cancer’s progression, treatment response, and long-term prognosis. Chemotherapy-induced lipid alterations may persist, increasing cardiovascular risk and potentially affecting therapeutic efficacy. Routine lipid monitoring and metabolic interventions could enhance treatment outcomes and survivorship. Future research should focus on developing lipid-targeted strategies to optimize breast cancer management. Full article

Physical stimulation, which is a key factor affecting the metabolism of osteoblasts and their precursor cells, plays an important role in bone remodeling; however, the role of micro-vibrations in osteoblast differentiation is unclear. In the present study, we determined the effects of frequency-regulated repeated micro-vibration (FRMV) on cell proliferation and established a method to induce osteoblast differentiation through FRMV using the mouse pre-osteoblast-like cell line MC3T3-E1, which is widely used in bone metabolism research. The results indicated that FRMV significantly influenced the proliferation of MC3T3-E1 cells in a normal growth medium. FRMV at 42.2 Hz significantly promoted proliferation, whereas FRMV at 92.1 Hz showed no effect on the proliferation rate. Moreover, FRMV at 42.2 Hz significantly increased alkaline phosphatase (ALP) enzyme activity and ALP gene expression in MC3T3-E1 cells. Treatment with LDN193189, a bone morphogenetic protein (BMP) signaling inhibitor, revealed that the FRMV-induced upregulation in ALP enzyme activity and ALP gene expression were significantly suppressed in MC3T3-E1 cells. The results suggest that the FRMV protocol developed in the present study induces osteoblast differentiation through the BMP signaling pathway. Thus, FRMV may contribute to the development of effective bone regeneration technologies. Full article

Introduction: Recombinant human collagen, developed through advanced recombinant DNA technology, has emerged as a cutting-edge biomaterial with diverse applications in medicine. It addresses significant limitations of animal-derived collagens, such as immunogenicity and the risk of zoonotic diseases. Objective: This review evaluates the clinical applications, benefits, and challenges associated with recombinant human collagen, focusing on its potential to transform medical and surgical practices. Methods: A comprehensive search was conducted in MEDLINE, PubMed, and Ovid databases using keywords such as “Recombinant Human Collagen”, “Collagen-Based Biomaterials”, “Clinical Applications”, “Tissue Repair”, and “Wound Healing”. Relevant studies, including clinical trials and diagnostic applications, were analyzed and classified according to the Oxford Centre for Evidence-Based Medicine evidence hierarchy. Findings: Recombinant human collagen demonstrates superior mechanical properties and controlled degradation rates compared to traditional collagen sources. Clinical studies highlight its effectiveness in accelerating wound closure, promoting dermal regeneration, and minimizing scarring, making it particularly valuable in chronic wound management and surgical interventions. In tissue engineering, recombinant human collagen scaffolds have shown potential for regenerating cartilage, bone, and cardiovascular tissues by supporting cell proliferation, differentiation, and matrix deposition. Additionally, its adaptability for forming hydrogels and matrices enhances its suitability for drug delivery systems, enabling controlled and sustained release of therapeutic agents. Conclusion: Recombinant human collagen represents a transformative advancement in clinical practice, providing a safer and more effective alternative to traditional collagen sources. Its demonstrated success in wound healing, tissue engineering, and drug delivery highlights its potential to significantly improve patient outcomes. However, challenges such as high production costs, regulatory complexities, and long-term biocompatibility remain barriers to widespread clinical adoption. Further research and collaboration between biotechnology developers and regulatory authorities are essential to fully realize its clinical potential. Full article

Osteoporosis can increase the risk of fracture in elderly patients, and insufficient control affects quality of life. Rikkunshi-To (RKT) has been prescribed for elderly patients to improve gastrointestinal function. We postulated that RKT has preventive potential for the development of osteoporosis. Thus, we developed a simple method to evaluate osteoporosis using a continuous series of X-ray images of femurs in mice, and investigated the effects of RKT on the development of osteoporosis in these mice. Male senescence-accelerated mouse strain P6 (SAMP6) mice, a model of senile osteoporosis in humans, were fed diets with or without RKT (1%). We collected X-ray images of the whole body of each mouse weekly and measured the ratio of cortical thickness of the femur (C/F index). The C/F index in SAMP6 mice fed the normal diet was increased between 50 and 80 days old, but it was significantly decreased after 120 days old. On the other hand, the C/F index in SAMP6 mice fed the RKT diet was increased between 50 and 80 days old; however, it remained unchanged throughout the experimental period. We also confirmed that the C/F index in SAMP6 mice fed the RKT diet suddenly decreased on the replacement of the RKT diet with a normal diet, suggesting that we can collect data related to a series of continuous changes in bone mass, and that RKT is useful for the prevention of osteoporosis. Full article

Toxic chemicals and epigenetic biomarkers associated with cancer have been used successfully in clinical diagnostic screening of feces and urine from individuals, but they have been underutilized in a global setting. We analyzed peer-reviewed literature to achieve the following: (i) compile epigenetic biomarkers of disease, (ii) explore whether research locations are geographically aligned with disease hotspots, and (iii) determine the potential for tracking disease-associated epigenetic biomarkers. Studies (n = 1145) of epigenetic biomarkers (n = 146) in urine and feces from individuals have established notable diagnostic potential for detecting and tracking primarily gastric and urinary cancers. Panels with the highest sensitivity and specificity reported more than once were SEPT9 (78% and 93%, respectively) and the binary biomarker combinations GDF15, TMEFF2, and VIM (93% and 95%), NDRG4 and BMP3 (98% and 90%), and TWIST1 and NID2 (76% and 79%). Screening for epigenetic biomarkers has focused on biospecimens from the U.S., Europe, and East Asia, whereas data are limited in regions with similar/higher disease incidence rates (i.e., data for New Zealand, Japan, and Australia for colorectal cancer). The epigenetic markers discussed here may aid in the future monitoring of multiple cancers from individual- to population-level scales by leveraging the emerging science of wastewater-based epidemiology (WBE). Full article

Objective: This study aims to assess the impact of prolonged occupational exposure to chrysotile asbestos on the epithelial cells of the upper respiratory tract and the levels of surfactant protein D (SP-D) in female workers. Methods: Buccal epithelial cell samples were collected from 40 workers at JSC “Kostanay Minerals”, fixed using the May–Grünwald method, and stained with the Romanowsky–Giemsa technique. SP-D levels were measured using an enzyme-linked immunosorbent assay (ELISA). Results: Workers exposed to asbestos dust exhibited a significant increase in cytological abnormalities and higher SP-D levels compared to the control group. Conclusion: Prolonged exposure to chrysotile-containing dust leads to degenerative changes in upper respiratory tract epithelial cells, characterized by cytological and cytogenetic abnormalities, alongside elevated SP-D levels, highlighting the need for preventive health measures. Full article

The efficacy of hematopoietic stem cell (HSC) therapy for cerebral infarction has been previously demonstrated. However, the lack of response in some patients has hindered its widespread use. To establish HSC therapy as a standard treatment, it is important to examine the causes of non-responsiveness. In this study, we aimed to identify the specifications of transplanted cells based on their therapeutic mechanisms to predict treatment success. We found that HSC therapy activates injured cerebral endothelial cells via gap junctions because cell adhesion between HSCs and the endothelium plays an essential role in cellular communication via gap junctions. The expression of the adhesion molecule integrin β2 (CD18) in CD34-positive (CD34⁺) cells was identified as critical for the therapeutic effect on cerebral infarction in a murine model. Cells with low CD18 expression exhibited a weaker therapeutic effect than cells with high CD18 expression, even when the same number of HSCs was administered. The expression of CD18 in CD34⁺ cells can be used as a specification marker for transplanted HSCs and is useful for identifying non-responders. Furthermore, quantification of CD18 expression is crucial for evaluating the cellular potential of cell-based therapies for diseases where therapeutic effects are mediated through cell adhesion. Full article

Resveratrol can beneficially affect growth and follicle development and lead to improved sperm function parameters in pre-clinical studies, while information from clinical studies is still inconclusive. This study aims to evaluate the biological and clinical impact of a resveratrol-based multivitamin supplement on level II assisted reproduction cycles (IVF and intracytoplasmic sperm injection [ICSI]). A retrospective, case-control study, involving 70 infertile couples undergoing IVF/ICSI cycles, was conducted at the Assisted Reproductive Center, Obstetrics and Gynecology Unit-Villa Sofia-Cervello Hospital in Palermo. The study group underwent pre-treatment with a daily nutraceutical based on resveratrol, whereas the control group received 400 mcg/day of folic acid. Primary endpoints to be evaluated were the number of mature follicles developed (>16 mm), total oocytes and Metaphase II (MII) oocytes retrieved, fertilization rate, number of embryos/blastocysts obtained, and semen quality. Secondary objectives in our evaluation were the duration and dosage of gonadotropins, the starting dose, the number of blastocysts to be transferred and frozen, implantation rate, and, ultimately, biochemical and clinical pregnancy rates. In the study group, a significantly higher number of mature follicles, oocytes, and MII oocytes were collected compared to the control group. In the study group, a higher fertilization rate as well as higher numbers of cleavage embryos per patient, blastocysts per patient, and frozen blastocysts were obtained. In the study group, a shorter administration time and lower dosages of gonadotropins required to reach follicle maturity were also observed compared to controls, with fewer dose adjustments during stimulation compared to the starting dose. No significant differences were found in biochemical or clinical pregnancy rates. A 12-month period of dietary supplementation with a resveratrol-based multivitamin nutraceutical leads to better biological effects on ICSI cycles. Full article

Parkinson’s disease (PD) is the second-leading cause of death among neurodegenerative disease after Alzheimer’s disease (AD), affecting around 2% of the population. It is expected that the incidence of PD will exceed 12 million by 2040. Meanwhile, there is a recognized difference in the phenotypical expression of the disease and response to treatment between men and women. Men have twice the incidence of PD compared to women, who have a late onset and worse prognosis that is usually associated with menopause. In addition, the incidence of PD in women is associated with the cumulative estrogen levels in their bodies. These differences are suggested to be due to the protective effect of estrogen on the brain, which cannot be given in clinical practice to improve the symptoms of the disease because of its peripheral side effects, causing cancer in both males and females in addition to the feminizing effect it has on males. As PD pathophysiology involves alteration in the expression levels of multiple LncRNAs, including metastatic-associated lung adenocarcinoma transcript 1 (MALAT1), and as estrogen has been illustrated to control the expression of MALAT1 in multiple conditions, it is worth investigating the estrogen–MALAT1 interaction in Parkinson’s disease to mimic its protective effect on the brain while avoiding its peripheral side effects. The following literature review suggests the potential regulation of MALAT1 by estrogen in PD, which would enhance our understanding of the pathophysiology of the disease, improving the development of more tailored and effective treatments. Full article

The human pineal gland is the largest producer of the hormone melatonin. Pineal acervuli (brain sand), calcified concretions in the pineal gland, have long been studied because of their association with ageing, melatonin production, and neurological disorders. The solid inorganic matter of the hydroxyapatite crystals often renders sample sectioning impossible, to the extent that the sections lose value. Technically, freeze-fracturing has revealed the detailed structure and cell relationships without tissue damage. In our electron microscopic study, samples of the human pineal gland were obtained during autopsy from 20 donors with mean age 69 years. Samples underwent freeze-fracturing and standard histological procedures, and were analysed by scanning electron microscopy (SEM) in high vacuum. Based on our results, freeze-fracturing enabled identification of a mulberry-like acervulus topography. The acervuli were situated in specific “nest-like” structures, which were surrounded by pinealocytes, interstitial cells, and nerve fibres. A fractured surface of the intrapineal acervuli exhibited a regular lamellar structure. Freeze-fracturing the pineal gland and imaging by SEM enabled complex structural analysis. This approach permits viewing the surface acervuli spherical and internal lamellar architecture. Our results confirmed that the parenchyma of this small but important gland contains two types of acervuli, depending on their size: non-aggregated and aggregated. We propose to include these forms of acervuli in the new edition of the Terminologica Histologica. In conclusion, pineal gland freeze-fracturing by SEM is suitable for complex structural analysis. Our description of our methods can be a guide for other scientists who want to study the pineal gland with electron microscopy methods. Full article

A CpG oligodeoxynucleotide (CpG-ODN), iSN40, was originally identified as promoting the mineralization and differentiation of osteoblasts, independent of Toll-like receptor 9 (TLR9). Since CpG ODNs are often recognized by TLR9 and inhibit osteoclastogenesis, this study investigated the TLR9 dependence and anti-osteoclastogenic effect of iSN40 to validate its potential as an osteoporosis drug. The murine monocyte/macrophage cell line RAW264.7 was treated with the receptor activator of nuclear factor-κB ligand (RANKL) to induce osteoclast differentiation, then the effect of iSN40 on was quantified by tartrate-resistant acid phosphatase (TRAP) staining and real-time RT-PCR. iSN40 completely inhibited RANKL-induced differentiation into TRAP⁺ multinucleated osteoclasts by suppressing osteoclastogenic genes and inducing anti-/non-osteoclastogenic genes. Treatment with a TLR9 inhibitor, E6446, or a mutation in the CpG motif of iSN40 abolished the intracellular uptake and anti-osteoclastogenic effect of iSN40. These results demonstrate that iSN40 is subcellularly internalized and is recognized by TLR9 via its CpG motif, modulates RANKL-dependent osteoclastogenic gene expression, and ultimately inhibits osteoclastogenesis. Finally, iSN40 was confirmed to inhibit the osteoclastogenesis of RAW264.7 cells cocultured with the murine osteoblast cell line MC3T3-E1, presenting a model of bone remodeling. This study demonstrates that iSN40, which exerts both pro-osteogenic and anti-osteoclastogenic effects, may be a promising nucleic acid drug for osteoporosis. Full article

Despite the promising potential of cell-based therapies developed using tissue engineering techniques to treat a wide range of diseases, including limbal stem cell deficiency (LSCD), which leads to corneal blindness, their commercialization remains constrained. This is primarily attributable to the limited cell sources, the use of non-standardizable, unscalable, and unsustainable techniques, and the extended manufacturing processes required to produce transplantable tissue-like surrogates. Herein, we present the first demonstration of the potential of a novel approach combining collagen films (CF), hyaluronic acid (HA), human telomerase-immortalized limbal epithelial stem cells (T-LESCs), and macromolecular crowding (MMC) to develop innovative biomimetic substrates for limbal epithelial stem cells (LESCs). The initial step involved the fabrication and characterization of CF and CF enriched with HA (CF-HA). Subsequently, T-LESCs were seeded on CF, CF-HA, and tissue culture plastic (TCP). Thereafter, the effect of these matrices on basic cellular function and tissue-specific extracellular matrix (ECM) deposition with or without MMC was evaluated. The viability and metabolic activity of cells cultured on CF, CF-HA, and TCP were found to be similar, while CF-HA induced the highest (p < 0.05) cell proliferation. It is notable that CF and HA induced cell growth, whereas MMC increased (p < 0.05) the deposition of collagen IV, fibronectin, and laminin in the T-LESC culture. The data highlight the potential of, in particular, immortalized cells and MMC for the development of biomimetic cell culture substrates, which could be utilized in ocular surface reconstruction following further in vitro, in vivo, and clinical validation of the approach. Full article