Proliferating and migrating effects of regenerating sea anemone Aulactinia stella cells-derived exosomes on human skin fibroblasts.

Regenerative effects of sea anemone-derived exosomes on human foreskin fibroblasts (HFFs) were investigated. Water-based extracts from regenerating Aulactinia stella tissue were collected at various time points, and exosomes were extracted after inducing wounds. Both the extract and exosomes were tested on HFF for proliferation and in vitro wound healing. In silico analysis explored protein-protein docking between regenerative exosome proteins and HFF receptors. The MTT (3-(4,5-dimethylthiazol-2yl)-2,5 diphenyltetrazolium bromide proliferation assay and in vitro wound healing test of aquatic extract showed proliferative effects on HFF cell lines, with the 60 µg/mL concentration significantly enhancing cell migration. Exosomes were characterised. Exosomes showed a significantly positive effect on cell proliferation and migration at the 50 µg/mL concentration 48 h post-wound induction. In silico analysis revealed potential binding affinities between exosome proteins and HFF receptors. In conclusion, optimised concentrations of A. stella-derived exosomes exhibited positive effects on HFF regeneration and migration, suggesting their potential in accelerating wound healing.

Tissue clearing and three-dimensional imaging of intact tissues: a review on FACT protocol.

FACT is a developed technique for clearing tissues that does not use acrylamide. Since the removal of lipids is crucial for transparency and efficient antibody staining throughout the tissue, especially for microscopy and imaging, it is a harmful process that can cause the loss of important biological molecules such as proteins. The FACT technique overcomes this by chemically bonding the membrane and intracellular proteins with the extracellular matrix, creating a massive 3D hydrogel matrix and providing structural support to fortify the tissue during processing. Compared to other acrylamide-based techniques, the FACT technique requires less labor and harmful chemicals and is therefore considered a more suitable option. In this study, we describe the complete FACT protocol for antibody staining and imaging of whole-cleared tissues while preserving structure and improving image quality. The protocol includes tissue perfusion, fixation, clearing, antibody staining, refractive index matching (RIM) (), microscopy, and imaging. The timing for each step varies depending on the size, weight, and type of tissue, as well as the type of immunostaining. We provide an example of the FACT protocol using mouse brain tissue, which demonstrates its suitability for molecular interrogation analysis of large tissues. The FACT technique has been successfully performed on different types of tissues, making it a favorable choice for a variety of applications.

Sex differentials in the prevalence of behavioral risk factors and non-communicable diseases in adult populations of West Kazakhstan.

Introduction: The prevalence of non-communicable diseases (NCDs) is increasing worldwide. Several modifiable risk factors, such as smoking, alcohol drinking, physical inactivity, and obesity, have been linked to the development of NCDs in both genders. Understanding the prevalence of these risk factors and their associated factors is crucial for effective intervention planning in adult populations. This study aimed to provide an overview of the prevalence and associated factors of these risk behaviors among different genders of adults in West Kazakhstan. Methods: A cross-sectional study was conducted in four regions of West Kazakhstan. A stratified multistage sampling technique was utilized to obtain a representative sample size of 4,800 participants aged 18 -69 years. Trained researchers administered face-to-face interviews using validated questionnaires to gather information pertaining to sociodemographic characteristics, smoking habits, alcohol drinking, dietary patterns, physical activity levels, body mass index (BMI), and prevalent diseases. Results: This study, which included 4,800 participants from West Kazakhstan, revealed some striking numerical findings. The overall prevalence rates of behavioral risk factors and metabolic conditions were as follows: smoking was 13.6% (95%CI: 3.2-24.0%), alcohol drinking was 47.0% (27.7-66.3%), current obesity was 22.3% (9.0-35.6%), and physical inactivity was 80.7% (55.4-106.0%). In addition, the overall prevalence rates of metabolic conditions were 25.6% (11.3-39.9%) for hypertension, 7.5% (0.2-15.2%) for diabetes, 11.8% (2.1-21.5%) for high cholesterol, and 13.0% (2.8-23.2%) for cardiovascular diseases. Additionally, a higher prevalence of high cholesterol was observed in men, and a greater prevalence of heart disease was identified in women. Multinomial logistic regression revealed that physical inactivity was associated with hypertension, diabetes, and heart disease, while obesity was linked to hypertension, high cholesterol, and heart disease. Discussion: This study in West Kazakhstan identified variations in the prevalence of behavioral risk factors and NCDs, highlighting gender, age, and regional disparities. Notably, men showed higher rates of smoking and alcohol drinking, while women exhibited a greater prevalence of physical inactivity and obesity. Gender and regional differences were evident, with the West Kazakhstan region standing out for distinct patterns. Tailored interventions are crucial to address these disparities and enhance public health in the region.

Enhancing differentiation of menstrual blood-derived stem cells into female germ cells using a bilayer amniotic membrane and nano-fibrous fibroin scaffold.

Three-dimensional nanofiber scaffolds offer a promising method for simulating in vivo conditions within the laboratory. This study aims to investigate the influence of a bilayer amniochorionic membrane/nanofibrous fibroin scaffold on the differentiation of human menstrual blood mesenchymal stromal/stem cells (MenSCs) into female germ cells. MenSCs were isolated and assigned to four culture groups: (i) MenSCs co-cultured with granulosa cells (GCs) using the scaffold (3D-T group), (ii) MenSCs using the scaffold alone (3D-C group), (iii) MenSCs co-cultured only with GCs (2D-T group), and (iv) MenSCs without co-culture or scaffold (2D-C group). Both MenSCs and GCs were independently cultured for two weeks before co-culturing was initiated. Flow cytometry was employed to characterize MenSCs based on positive markers (CD73, CD90, and CD105) and negative markers (CD45 and CD133). Additionally, flow cytometry and immunocytochemistry were used to characterize the GCs. Differentiated MenSCs were analyzed using real-time PCR and immunostaining. The real-time PCR results demonstrated significantly higher levels of VASA expression in the 3D-T group compared to the 3D-C, 2D-T, and 2D-C groups. Similarly, the SCP3 mRNA level in the 3D-T group was notably elevated compared to the 3D-C, 2D-T, and 2D-C groups. Moreover, the expression of GDF9 was significantly higher in the 3D-T group when compared to the 3D-C, 2D-T, and 2D-C groups. Immunostaining results revealed a lack of signal for VASA, SCP3, or GDF9 markers in the 2D-T group, while some cells in the 3D-T group exhibited positive staining for all these proteins. These findings suggest that the combination of a bilayer amniochorionic membrane/nanofibrous fibroin scaffold with co-culturing GCs facilitates the differentiation of MenSCs into female germ cells.