Background: Cells are sensitive to changes in gravity, especially the cytoskeletal structures that determine cell morphology. The aim of this study was to assess the effects of simulated microgravity (SMG) on 3T3 cell morphology, as demonstrated by a characterization of the morphology of cells and nuclei, alterations of microfilaments and microtubules, and changes in cycle progression. Methods: 3T3 cells underwent induced SMG for 72 h with Gravite^®, while the control group was under 1G. Fluorescent staining was applied to estimate the morphology of cells and nuclei and the cytoskeleton distribution of 3T3 cells. Cell cycle progression was assessed by using the cell cycle app of the Cytell microscope, and Western blot was conducted to determine the expression of the major structural proteins and main cell cycle regulators. Results: The results show that SMG led to decreased nuclear intensity, nuclear area, and nuclear shape and increased cell diameter in 3T3 cells. The 3T3 cells in the SMG group appeared to have a flat form and diminished microvillus formation, while cells in the control group displayed an apical shape and abundant microvilli. The 3T3 cells under SMG exhibited microtubule distribution surrounding the nucleus, compared to the perinuclear accumulation in control cells. Irregular forms of the contractile ring and polar spindle were observed in 3T3 cells under SMG. The changes in cytoskeleton structure were caused by alterations in the expression of major cytoskeletal proteins, including β-actin and α-tubulin 3. Moreover, SMG induced 3T3 cells into the arrest phase by reducing main cell cycle related genes, which also affected the formation of cytoskeleton structures such as microfilaments and microtubules. Conclusions: These results reveal that SMG generated morphological changes in 3T3 cells by remodeling the cytoskeleton structure and downregulating major structural proteins and cell cycle regulators.

This study aimed to assess the effects of hexavalent chromium on zebrafish (Danio rerio) embryo development. The zebrafish embryos were treated with solutions containing chromium at different concentrations (0.1, 1, 3.125, 6.25, 12.5, 50, and 100 µg/mL). The development of zebrafish embryos was estimated by the determination of survival rate, heart rate, and the measurement of larvae body length. Real time RT-PCR and Western blot were performed to assess the expression of apoptosis- and antioxidant-related genes. The results showed that the reduced survival rate of zebrafish embryos and larvae was associated with an increase in chromium concentration. The exposure of higher concentrations resulted in a decrease in body length of zebrafish larvae. In addition, a marked increase in heart rate was observed in the zebrafish larvae under chromium treatment, especially at high concentrations. The real-time RT-PCR analysis showed that the transcript expressions for cell-cycle-related genes (cdk4 and cdk6) and antioxidant-related genes (sod1 and sod2) were downregulated in the zebrafish embryos treated with chromium. Western blot analysis revealed the upregulation of Caspase 3 and Bax, while a downregulation was observed in Bcl2. These results indicated that hexavalent chromium induced changes in zebrafish embryo development by altering apoptosis- and antioxidant-related genes.

Dermal Papilla Cells (DPCs) are mesenchymal cells residing at the base of the hair follicles. They play an important role in the hair cycle remodeling and hair follicle regeneration. In this study, DPCs were isolated, and cultured from human hair follicles and evaluated the expression of -SMA, versican by immunocytochemistry, and alkaline phosphatase activity. The human fibroblasts were used as a control to confirm the non-fibroblast DPCs. The results showed that the cells grew from the hair bulb and had fibroblast morphology. These bulb cells strongly expressed alkaline phosphatase activity -SMA, and versican while the human fibroblast lacked ALP activity and less expression of -SMA and versican. In conclusion, the protocol is established to isolate and culture DPCs from hair follicle bulbs, and ALP activity is suggested as the major marker for distinguishing DPCs and human fibroblasts.

Drug strategy is a standard method for treating type 2 diabetes mellitus (T2D), a non-communicable disease with increasing prevalence, which may cause side effects. Therefore, natural compounds with limited adverse effects have come back into vogue for treating T2D. This study aims to evaluate the effects on rehabilitating hyperglycemic mice of cashew nut testa (husk) extract and fraction known as potential bio-substances for improvement in T2D. First, the hyperglycemic mice were induced with a high-fat diet (HFD) for 4 weeks and then were injected with streptozotocin (STZ, dozen for injection was 40 mg/kg/week) for 2 weeks. Next, the confirmed hyperglycemic mice were treated with pioglitazone (HG+PG group), total extract (HG+TE group), and saponin-rich fraction (HG+SRF group) for 3 weeks. Then, the evaluation was based on body mass; blood glucose (BG) level; BG tolerance, lipid profile, pancreatic histology and the expression IRS-1 in the pancreas. The results showed that body mass and BG level significantly increased in hyperglycemic mice. After substance treatment, there was no change in body mass in TE and SRF groups. However, BG level of HG+TE group mice significantly decreased compared to hyperglycemic mice and only BG tolerance of HG+SRF group was improved. Besides, HG+TE and HG+SRF groups modulated the triglyceride, HDL and LDL close to those expressed in normal mice. In addition, histological images of the pancreas revealed the restoration in both HG+TE and HG+SRF groups. Simultaneously, the IRS-1 expression in HG+TE group pancreas was restored to its expression in normal mice. These results demonstrate that the TE and SRF of cashew nut testa could ameliorate BG, lipid profile and pancreatic IRS-1 expression and restore the damaged pancreas and islets in hyperglycemic mice.

Introduction: Mesenchymal stem cells (MSCs) are possibly the most potent type of stem cells for the treatment of many diseases since they possess many advantageous properties, such as abundant source, ease of isolation, and potential to differentiate and trans-differentiate into different types of cells. Although the therapeutic potential of expanded MSCs has been well proven, their biosafety features have not been fully understood. This study aimed to investigate some changes in phenotype and gene expression of bone marrow derived MSCs after long term expansion. Methods: In this study, expanded mesenchymal stem cells derived from human bone marrow (hBMSCs) were identified for their characteristics (which included morphology, immunophenotype, and differentiation potential) at passages 5, 10 and 15. Moreover, they were evaluated for the expression of various tumor suppressor genes (PTEN, p16, and p53) by real-time RT-PCR. Results: The results showed that the hBMSCs at passage 15 displayed a change in morphology and a slight reduction of the expression of CD44 and CD90, whereas their potential for adipogenic and osteogenic differentiation was maintained. Moreover, the expression of tumor suppressor genes in the hBMSCs increased after long-term culture. Conclusion: It could be assumed that prolonged cultures of more than 15 passages drove the hBMSCs into senescence phase. Cultured hBMSCs below passage 10 seemed to be more effective in application because their properties were still preserved.