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Objective: Disruptor of telomeric silencing 1-like (DOT1L) is the only known histone H3K79 methyltransferase essential for the development of the embryonic cardiovascular system, including the heart, blood vessels, and lymphatic vessels, through transcriptional regulation. Our previous study demonstrated that Dot1l deletion results in aberrant lymphatic development and function. However, its precise function in the postnatal cardiovascular system remains unknown. Methods: Using conditional and inducible Dot1l knockout (KO) mice, along with a reporter strain carrying the Geo gene at the Dot1l locus, DOT1L expression and its function in the vascular system during postnatal life were investigated. To assess vessel morphology and vascular permeability, we administered Latex or Evans blue dye to KO mice. In addition, in vitro tube formation and cell migration assays were performed using DOT1L-depleted human umbilical vein endothelial cells (HUVECs). Changes in the expression of vascular genes in HUVECs were measured by quantitative polymerase chain reaction. Results: Our findings demonstrate that conditional Dot1l knockout in the Tg (Tie2-cre) strain results in abnormal blood vessel formation and lymphatic anomalies in the intestine. In a mouse model of Rosa26-creER-mediated inducible Dot1l knockout, we observed vascular phenotypes, including increased vascular permeability and brain hemorrhage, when DOT1L was deleted in adulthood. Additionally, DOT1L depletion in cultured HUVECs led to impaired cell migration and tube formation, likely due to altered gene transcription. These findings highlight the essential role of DOT1L in maintaining vascular integrity and function during embryonic development and postnatal life. Conclusion: Our study revealed that DOT1L is required for the maintenance of adult vascular function through the regulation of gene expression. [ABSTRACT FROM AUTHOR]

Objective: R-loops are DNA:RNA triplex hybrids, and their metabolism is tightly regulated by transcriptional regulation, DNA damage response, and chromatin structure dynamics. R-loop homeostasis is dynamically regulated and closely associated with gene transcription in mouse zygotes. However, the factors responsible for regulating these dynamic changes in the R-loops of fertilized mouse eggs have not yet been investigated. This study examined the functions of candidate factors that interact with R-loops during zygotic gene activation. Methods: In this study, we used publicly available next-generation sequencing datasets, including low-input ribosome profiling analysis and polymerase II chromatin immunoprecipitation-sequencing (ChIP-seq), to identify potential regulators of R-loop dynamics in zygotes. These datasets were downloaded, reanalyzed, and compared with mass spectrometry data to identify candidate factors involved in regulating R-loop dynamics. To validate the functions of these candidate factors, we treated mouse zygotes with chemical inhibitors using in vitro fertilization. Immunofluorescence with an anti-R-loop antibody was then performed to quantify changes in R-loop metabolism. Results: We identified DEAD-box-5 (DDX5) and histone deacetylase-2 (HDAC2) as candidates that potentially regulate R-loop metabolism in oocytes, zygotes and two-cell embryos based on change of their gene translation. Our analysis revealed that the DDX5 inhibition of activity led to decreased R-loop accumulation in pronuclei, indicating its involvement in regulating R-loop dynamics. However, the inhibition of histone deacetylase-2 activity did not significantly affect R-loop levels in pronuclei. Conclusion: These findings suggest that dynamic changes in R-loops during mouse zygote development are likely regulated by RNA helicases, particularly DDX5, in conjunction with transcriptional processes. Our study provides compelling evidence for the involvement of these factors in regulating R-loop dynamics during early embryonic development. [ABSTRACT FROM AUTHOR]

Over the past few decades, carbon nanomaterials, including carbon nanofibers, nanocrystalline diamonds, fullerenes, carbon nanotubes, carbon nanodots, and graphene and its derivatives, have gained the attention of bioengineers and medical researchers as they possess extraordinary physicochemical, mechanical, thermal, and electrical properties. Recently, surface functionalization with carbon nanomaterials in dental and orthopedic implants has emerged as a novel strategy for reinforcement and as a bioactive cue due to their potential for osseointegration. Numerous developments in fabrication and biological studies of carbon nanostructures have provided various novel opportunities to expand their application to hard tissue regeneration and restoration. In this minireview, the recent research trends in surface functionalization of orthopedic and dental implants with coating carbon nanomaterials are summarized. In addition, some seminal methodologies for physicomechanical and electrochemical coatings are discussed. In conclusion, it is shown that further development of surface functionalization with carbon nanomaterials may provide innovative results with clinical potential for improved osseointegration after implantation. [ABSTRACT FROM AUTHOR]

Tissue engineering has recently emerged as a novel strategy for the regeneration of damaged skeletal muscle tissues due to its ability to regenerate tissue. However, tissue engineering is challenging due to the need for state-of-the-art interdisciplinary studies involving material science, biochemistry, and mechanical engineering. For this reason, electrospinning and three-dimensional (3D) printing methods have been widely studied because they can insert embedded muscle cells into an extracellular-matrix-mimicking microenvironment, which helps the growth of seeded or laden cells and cell signals by modulating cell–cell interaction and cell–matrix interaction. In this mini review, the recent research trends in scaffold fabrication for skeletal muscle tissue regeneration using advanced techniques, such as electrospinning and 3D bioprinting, are summarized. In conclusion, the further development of skeletal muscle tissue engineering techniques may provide innovative results with clinical potential for skeletal muscle regeneration. [ABSTRACT FROM AUTHOR]

Recent studies have demonstrated the generation of midbrain-like organoids (MOs) from human pluripotent stem cells. However, the low efficiency of MO generation and the relatively immature and heterogeneous structures of the MOs hinder the translation of these organoids from the bench to the clinic. Here we describe the robust generation of MOs with homogeneous distribution of midbrain dopaminergic (mDA) neurons. Our MOs contain not only mDA neurons but also other neuronal subtypes as well as functional glial cells, including astrocytes and oligodendrocytes. Furthermore, our MOs exhibit mDA neuron-specific cell death upon treatment with 1-methyl-4-phenyl-1,- 2,3,6-tetrahydropyridine, indicating that MOs could be a proper human model system for studying the in vivo pathology of Parkinson's disease (PD). Our optimized conditions for producing homogeneous and mature MOs might provide an advanced patientspecific platform for in vitro disease modeling as well as for drug screening for PD. [ABSTRACT FROM AUTHOR]

Peroxisome proliferator-activated receptor (PPAR) δ plays a pivotal role in metabolic homeostasis through its effect on insulin signaling. Although diverse genomic actions of PPARδ are postulated, the specific molecular mechanisms whereby PPARδ controls insulin signaling have not been fully elucidated. We demonstrate here that short-term activation of PPARδ results in the formation of a stable complex with nuclear T-cell protein tyrosine phosphatase 45 (TCPTP45) isoform. This interaction of PPARδ with TCPTP45 blocked translocation of TCPTP45 into the cytoplasm, thereby preventing its interaction with the insulin receptor, which inhibits insulin signaling. Interaction of PPARδ with TCPTP45 blunted interleukin 6-induced insulin resistance, leading to retention of TCPTP45 in the nucleus, thereby facilitating deactivation of the signal transducer and activator of transcription 3 (STAT3)-suppressor of cytokine signaling 3 (SOCS3) signal. Finally, GW501516-activated PPARδ improved insulin signaling and glucose intolerance in mice fed a high-fat diet through its interaction with TCPTP45. This novel interaction of PPARδ constitutes the most upstream component identified of the mechanism downregulating insulin signaling. [ABSTRACT FROM AUTHOR]

The leading reason for implant revision surgery globally is lack of implant integration with surrounding bone. A new titanium alloy GUMMETAL® (Ti59Nb36Ta2Zr3O0.3) is currently used in biomedical devices and has a Young’s modulus that is better matched to bone. The surface was subject to NaOH, CaCl2, heat and water treatment (BioGum) after which the surfaces were evaluated using atomic force microscope, scanning electron microscope, X-ray diffractometer and elemental analysis using energy dispersive X-ray. To demonstrate enhanced bone bonding ability and cytocompatibility, apatite formation in simulated body fluid and in vitro stem cell attachment, proliferation and cytoskeleton organisation were examined. The formation of a ~200 nm nanoscale needle-like calcium titanate network on the surface following treatment was revealed and upon soaking in simulated body fluid, the formation of a ~5µm layer of apatite. Metabolic activity of rat bone marrow stem cells on BioGum was increased in comparison to control and the cell number appeared greater, with more elongated morphology as early as 2h post-seeding. This positions the modification as a simple and potentially universal technology for the improvement of implant integration. [ABSTRACT FROM AUTHOR]

In recent years, paramagnetic nanoparticles (NPs) have been widely used for magnetic resonance imaging (MRI). This paper reports the fabrication and toxicity evaluation of polyethylene glycol (PEG)-functionalized holmium oxide (Ho2O3) NPs for potential T2-weighted MRI applications. Various characterization techniques were used to examine the morphology, structure and chemical properties of the prepared PEG-Ho2O3 NPs. MRI relaxivity measurements revealed that PEG-Ho2O3 NPs could generate a strong negative contrast in T2-weighted MRI. The pilot cytotoxicity experiments showed that the prepared PEG-Ho2O3 NPs are biocompatible at concentrations less than 16 μg/mL. Overall, the prepared PEG-Ho2O3 NPs have potential applications for T2-weighted MRI imaging. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the effect of biphasic calcium phosphate (BCP) coated with reduced graphene oxide (rGO) as bone graft materials on bone regeneration. The rGO-coated BCP bone graft material was fabricatied by mixing rGO and BCP at various concentrations. The surface charge of rGO-coated BCP was measured to be -14.43 mV, which formed a static electrostatic interaction. Cell viabilities were significantly diminished at higher concentrations of ≥100 μg/mL. The calvarial defects of 48 rats were implanted rGO-coated BCPs at a weight ratio of 2:1000 (rGO2), 4:1000 (rGO4), and 10:1000 (rGO10), repectively. BCP was used as a control group. The micro-CT and histological analysis were performed to evaluate new bone formation at 2 and 8 weeks after surgery. The results showed that the new bone volume (mm3) was significantly higher in the experimental groups than in the control group. Histological analysis showed that new bone areas (%) were significantly higher in the rGO2 and rGO10 than in the control, and significantly higher in rGO4 than in the rGO2 and rGO10. Conclusively, the rGO-coated BCP was found to be effective on osteogenesis and the concentration of the composite was an important factor. [ABSTRACT FROM AUTHOR]

Nanoprobes with multimodal functionality have attracted significant interest recently because of their potential applications in nanomedicine. This paper reports the successful development of lanthanide-doped Y2O3 nanoprobes for potential applications in optical and magnetic resonance (MR) imaging. The morphology, structural, and optical properties of these nanoprobes were characterized by transmission electron microscope (TEM), field emission scanning electron microscope (FESEM), X-ray diffraction (XRD), energy-dispersive X-ray (EDX), and photoluminescence (PL). The cytotoxicity test showed that the prepared lanthanide-doped Y2O3 nanoprobes have good biocompatibility. The obvious contrast enhancement in the T1-weighted MR images suggested that these nanoprobes can be used as a positive contrast agent in MRI. In addition, the clear fluorescence images of the L-929 cells incubated with the nanoprobes highlight their potential for optical imaging. Overall, these results suggest that prepared lanthanide-doped Y2O3 nanoprobes can be used for simultaneous optical and MR imaging. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is a chronic, neurodegenerative disorder that results from the loss of cells in the substantia nigra (SN) which is located in the midbrain. However, no cure is available for PD. Recently, fibroblasts have been directly converted into induced neural stem cells (iNSCs) via the forced expression of specific transcription factors. Therapeutic potential of iNSC in PD has not been investigated yet. Here, we show that iNSCs directly converted from mouse fibroblasts enhanced functional recovery in an animal model of PD. The rotational behavior test was performed to assess recovery. Our results indicate that iNSC transplantation into the striatum of 6-hydroxydopamine (6-OHDA)-injected mice can significantly reduce apomorphine-induced rotational asymmetry. The engrafted iNSCs were able to survive in the striatum and migrated around the medial forebrain bundle and the SN pars compacta. Moreover, iNSCs differentiated into all neuronal lineages. In particular, the transplanted iNSCs that committed to the glial lineage were significantly increased in the striatum of 6-OHDA-injected mice. Engrafted iNSCs differentiated to dopaminergic (DA) neurons and migrated into the SN in the 6-OHDA lesion mice. Therefore, iNSC transplantation serves as a valuable tool to enhance the functional recovery in PD. [ABSTRACT FROM AUTHOR]

The optimum conditions for the fabrication of zein/Ag composite nanoparticles from ethanol/H2O cosolvents using electrospinning and the properties of the composite were investigated. The zein/Ag nanoparticles were characterized using field-emission scanning electron microscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD), and thermogravimetric analysis. The antibacterial activity of the zein/Ag composite nanoparticles was also investigated. The XRD patterns and TEM images indicate the coexistence of a zein matrix and well-distributed Ag nanoparticles. [ABSTRACT FROM AUTHOR]

Photolithography is a unique process that can effectively manufacture micro/nano-sized patterns on various substrates. On the other hand, the meniscus-dragging deposition (MDD) process can produce a uniform surface of the substrate. Graphene oxide (GO) is the oxidized form of graphene that has high hydrophilicity and protein absorption. It is widely used in biomedical fields such as drug delivery, regenerative medicine, and tissue engineering. Herein, we fabricated uniform GO micropatterns via MDD and photolithography. The physicochemical properties of the GO micropatterns were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), and Raman spectroscopy. Furthermore, cell migration on the GO micropatterns was investigated, and the difference in cell migration on triangle and square GO micropatterns was examined for their effects on cell migration. Our results demonstrated that the GO micropatterns with a desired shape can be finely fabricated via MDD and photolithography. Moreover, it was revealed that the shape of GO micropatterns plays a crucial role in cell migration distance, speed, and directionality. Therefore, our findings suggest that the GO micropatterns can serve as a promising biofunctional platform and cell-guiding substrate for applications to bioelectric devices, cell-on-a-chip, and tissue engineering scaffolds. [ABSTRACT FROM AUTHOR]

Human adipose-derived stem cells (hASCs) have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation. Recently, hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time. Matrix metalloproteinases (MMPs) are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells. MMPs secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs. [ABSTRACT FROM AUTHOR]

Background: In the field of biomedical engineering, many studies have focused on the possible applications of graphene and related nanomaterials due to their potential for use as scaffolds, coating materials and delivery carriers. On the other hand, electrospun nanofiber matrices composed of diverse biocompatible polymers have attracted tremendous attention for tissue engineering and regenerative medicine. However, their combination is intriguing and still challenging. Results: In the present study, we fabricated nanofiber matrices composed of M13 bacteriophage with RGD peptide displayed on its surface (RGD-M13 phage) and poly(lactic-co-glycolic acid, PLGA) and characterized their physicochemical properties. In addition, the effect of graphene oxide (GO) on the cellular behaviors of C2C12 myoblasts, which were cultured on PLGA decorated with RGD-M13 phage (RGD/PLGA) nanofiber matrices, was investigated. Our results revealed that the RGD/PLGA nanofiber matrices have suitable physicochemical properties as a tissue engineering scaffold and the growth of C2C12 myoblasts were significantly enhanced on the matrices. Moreover, the myogenic differentiation of C2C12 myoblasts was substantially stimulated when they were cultured on the RGD/PLGA matrices in the presence of GO. Conclusion: In conclusion, these findings propose that the combination of RGD/PLGA nanofiber matrices and GO can be used as a promising strategy for skeletal tissue engineering and regeneration. [ABSTRACT FROM AUTHOR]

This study investigated the effects of a specific regimen of resistance training on coordinated actions of human hand digits during grasping. Participants were instructed to hold a rectangular object with all five digits and to maintain the orientation of the object against transient perturbation. Indices of co-varied actions (i.e., synergies) among multi-digit grasping and rotational actions were quantified. The index of anticipatory changes of co-varied actions among digit forces (i.e., anticipatory synergy adjustment) was also quantified, which represents the controller's ability to predict an upcoming perturbation. The synergies of both grasping force and moment stabilization increased with the training. No change in the index of anticipatory synergy adjustment with training was observed. The current results suggest that the resistance training on the wrist could be an effective way to enhance both voluntary muscle force / torque production capability and ability to stabilize task performances during multi-digit prehensile tasks. [ABSTRACT FROM AUTHOR]

In biomedical applications, there is a need for tissue engineering scaffolds to promote and control cellular behaviors, including adhesion, proliferation and differentiation. In particular, the initial adhesion of cells has a great influence on those cellular behaviors. In this study, we concentrate on developing cell-adhesive substrates applicable for tissue engineering scaffolds. The hybrid nanofiber sheets were prepared by electrospinning poly(lactic-co-glycolic acid) (PLGA) and M13 phage, which was genetically modified to enhance cell adhesion thru expressing RGD peptides on their surface. The RGD peptide is a specific motif of extracellular matrix (ECM) for integrin receptors of cells. RGD peptide-decorated PLGA (RGD-PLGA) nanofiber sheets were characterized by scanning electron microscopy, immunofluorescence staining, contact angle measurement and differential scanning calorimetry. In addition, the initial adhesion and proliferation of four different types of mammalian cells were determined in order to evaluate the potential of RGD-PLGA nanofiber sheets as cell-adhesive substrates. Our results showed that the hybrid nanofiber sheets have a three-dimensional porous structure comparable to the native ECM. Furthermore, the initial adhesion and proliferation of cells were significantly enhanced on RGD-PLGA sheets. These results suggest that biomimetic RGD-PLGA nanofiber sheets can be promising cell-adhesive substrates for application as tissue engineering scaffolds. [ABSTRACT FROM AUTHOR]

Background: Electrospinning is a simple and effective method for fabricating micro- and nanofiber matrices. Electrospun fibre matrices have numerous advantages for use as tissue engineering scaffolds, such as high surface area-to-volume ratio, mass production capability and structural similarity to the natural extracellular matrix (ECM). Therefore, electrospun matrices, which are composed of biocompatible polymers and various biomaterials, have been developed as biomimetic scaffolds for the tissue engineering applications. In particular, graphene oxide (GO) has recently been considered as a novel biomaterial for skeletal muscle regeneration because it can promote the growth and differentiation of myoblasts. Therefore, the aim of the present study was to fabricate the hybrid fibre matrices that stimulate myoblasts differentiation for skeletal muscle regeneration. Results: Hybrid fibre matrices composed of poly(lactic-co-glycolic acid, PLGA) and collagen (Col) impregnated with GO (GO-PLGA-Col) were successfully fabricated using an electrospinning process. Our results indicated that the GO-PLGA-Col hybrid matrices were comprised of randomly-oriented continuous fibres with a three-dimensional non-woven porous structure. Compositional analysis showed that GO was dispersed uniformly throughout the GO-PLGA-Col matrices. In addition, the hydrophilicity of the fabricated matrices was significantly increased by blending with a small amount of Col and GO. The attachment and proliferation of the C2C12 skeletal myoblasts were significantly enhanced on the GO-PLGA-Col hybrid matrices. Furthermore, the GO-PLGA-Col matrices stimulated the myogenic differentiation of C2C12 skeletal myoblasts, which was enhanced further under the culture conditions of the differentiation media. Conclusions: Taking our findings into consideration, it is suggested that the GO-PLGA-Col hybrid fibre matrices can be exploited as potential biomimetic scaffolds for skeletal tissue engineering and regeneration because these GO-impregnated hybrid matrices have potent effects on the induction of spontaneous myogenesis and exhibit superior bioactivity and biocompatibility. [ABSTRACT FROM AUTHOR]

Recently, many nanomedical studies have been focused on magnetic nanoparticles (MNPs) because MNPs possess attractive properties for potential uses in imaging, drug delivery, and theranostics. MNPs must have optimized size as well as functionalized surface for such applications. However, careful cytotoxicity and genotoxicity assessments to ensure the biocompatibility and biosafety of MNPs are essential. In this study, Fe3O4 MNPs of different sizes (approximately 10 and 100-150 nm) were prepared with different functional groups, hydroxyl (-OH) and amine (-NH2) groups, by coating their surfaces with tetraethyl orthosilicate (TEOS), 3-aminopropyltrimethoxysilane (APTMS) or TEOS/APTMS. Differential cellular responses to those surface-functionalized MNPs were investigated in normal fibroblasts vs. fibrosarcoma cells. Following the characterization of MNP properties according to size, surface charge and functional groups, cellular responses to MNPs in normal fibroblasts and fibrosarcoma cells were determined by quantifying metabolic activity, membrane integrity, and DNA stability. While all MNPs induced just about 5% or less cytotoxicity and genotoxicity in fibrosarcoma cells at lower than 500 μg/mL, APTMS-coated MNPs resulted in greater than 10% toxicity against normal cells. Particularly, the genotoxicity of MNPs was dependent on their dose, size and surface charge, showing that positively charged (APTMS- or TEOS/APTMS-coated) MNPs induced appreciable DNA aberrations irrespective of cell type. Resultantly, smaller and positively charged (APTMS-coated) MNPs led to more severe toxicity in normal cells than their cancer counterparts. Although it was difficult to fully differentiate cellular responses to various MNPs between normal fibroblasts and their cancer counterparts, normal cells were shown to be more vulnerable to internalized MNPs than cancer cells. Our results suggest that functional groups and sizes of MNPs are critical determinants of degrees of cytotoxicity and genotoxicity, and potential mechanisms of toxicity. [ABSTRACT FROM AUTHOR]

Multifunctional mesoporous silica nanocomposites are attractive carriers for targeted drug delivery in nanomedicine. Although promising developments have been made in the fabrication of multifunctional mesoporous silica nanocomposites, the design and mass production of novel multifunctional carriers are still challenging. This paper reports the facile one-pot fabrication of a multifunctional inorganic composite composed of superparamagnetic Fe3O4 nanoparticles and coated dye-functionalized mesoporous silica with a high specific surface area. The resulting composite particles had a tunable particle size, special open pore channels with high specific surface area, which is quite favorable for drug loading and release properties, as well as luminescent and superparamagnetic properties suitable for targeted drug delivery and tracking. This composite exhibited low toxicity, suggesting potential biomedical applications. [ABSTRACT FROM AUTHOR]

Nanostructured electrodes have recently received great attention as components in lithium rechargeable batteries, especially because of the high power produced by the fast kinetic properties of these unique structures. Here, we report the successful synthesis of various nanostructured morphologies of spinel lithium manganese oxide electrodes (nanorod, nanothorn sphere, and sphere) from a similarly shaped manganese dioxide precursor that was controlled with different aluminium contents by the hydrothermal method. Among these structures, nanothorn sphere structured LiAl0.02Mn1.98O4produces the highest discharge capacity of 129.8 mA h g−1, excellent rate capability (94.6 mA h g−1at 20 C, 72% of 0.2 C-rate discharge capacity) and stable cyclic retention for 50 cycles. The excellent kinetic properties of the nanothorn sphere structure are not only due to the nanothorn sphere electrode having high surface area but also because the critical amount of Al in the nanothorn sphere electrode was located at the Mn site (16d) instead of the Li site (8a). [ABSTRACT FROM AUTHOR]

Green chemical synthesis of Au nanoparticles (NPs) has been of great interest because of its potential biomedical applications. In this study, we successfully produced phytochemical-induced Au NPs cofunctionalized with gallic acid, protocatechuic acid, and isoflavone. They have a strong antioxidant effect and serve as effective reducing agents, inducing the immediate passivation of Au NPs. The properties of these green chemical Au NPs were characterized by TEM, UV/Vis and FT-IR spectroscopy, and ζ-potential measurements, and the Au NPs exhibited excellent homogeneity with an average diameter of 20 nm and high dispersity at all pH ranges, with long-term stability as well as excellent cytocompatibility. Molecular dynamics (MD) simulations were also carried out in order to reveal the surface stability of the Au NPs. The computational results indicate that there are strong interactions between the phytochemicals and Au NPs, especially in the Au/protocatechuic acid–isoflavone model. Phytochemical-stabilized Au NPs allowed about 40% H2O2to be removed at an NP concentration of 50 μg mL−1; this removal rate is the same as that achieved by 3000 units per mg catalase. Therefore, this novel synthesis route for Au NPs using phytochemical reducing agents may be effectively exploited for nonthermal-assisted reactions and one-pot processes of biological applications. [ABSTRACT FROM AUTHOR]

Epiblast stem cells (EpiSCs) derived from epiblast tissue of post-implantation embryos are pluripotent and can give rise to all three germ layers in teratoma assays. Introduction of the four transcription factors Oct4, Sox2, Klf4 and c-Myc into somatic cells has been shown to generate induced pluripotent stem cells (iPSCs) that are very similar to embryonic stem cells (ESCs) in a number of characteristics. However, generation of EpiSCs by the direct reprogramming of somatic cells using these transcription factors has not been shown to date. Here, we show that these transcription factors can be used to directly generate induced EpiSCs (iEpiSCs) under EpiSC culture conditions. iEpiSCs resemble EpiSCs in morphology, gene expression pattern, epigenetic status and chimaera-forming capability. This study demonstrates that the culture environment in transcription factor-mediated reprogramming determines the cell fate of the reprogrammed cell. We therefore hypothesize that it will eventually be possible to shape the identity of a directly reprogrammed cell simply by modulating culture conditions. [ABSTRACT FROM AUTHOR]

In order to prevent restenosis after angioplasty or stenting, one of the most popular targets is suppression of the abnormal growth and excess migration of vascular smooth muscle cells (VSMCs) with drugs. However, the drugs also adversely affect vascular endothelial cells (VECs), leading to the induction of late thrombosis. We have investigated the effect of epigallocatechin-3-gallate (EGCG) on the proliferation and migration of VECs and VSMCs. Both cells showed dose-dependent decrease of viability in response to EGCG while they have different IC50 values of EGCG (VECs, 150 μM and VSMCs, 1050 μM). Incubating both cells with EGCG resulted in significant reduction in cell proliferation irrespective of cell type. The proliferation of VECs were greater affected than that of VSMCs at the same concentrations of EGCG. EGCG exerted differential migration-inhibitory activity in VECs vs. VSMCs. The migration of VECs was not attenuated by 200 μM EGCG, but that of VSMCs was significantly inhibited at the same concentration of EGCG. It is suggested that that EGCG can be effectively used as an efficient drug for vascular diseases or stents due to its selective activity, completely suppressing the proliferation and migration of VSMCs, but not adversely affecting VECs migration in blood vessels. [ABSTRACT FROM AUTHOR]

Somatic cells can achieve a pluripotent cell state in a process called pluripotential reprogramming. Multipotent stem cells can differentiate into cells of only one lineage, but pluripotent stem cells can give rise to cells of all three germ layers of an organism. In this study, we generated induced pluripotent stem (iPS) cells from bimaternal (uniparental) parthenogenetic neural stem cells (pNSCs) by transduction with either four (4F: Oct4, Klf4, Sox2, and c-Myc) or two (2F: Oct4 and Klf4) transcription factors. The resultant maternal iPS cells, which were reprogrammed directly from pNSCs, were capable of generating germ line-competent chimeras. Interestingly, analysis of global gene expression and imprinting status revealed that parthenogenetic iPS cells clustered closer to parthenogenetic ESCs than to female ESCs, with patterns that were clearly distinct from those of pNSCs. [ABSTRACT FROM AUTHOR]

Nestin is an intermediate filament protein expressed specifically in neural stem cells and progenitor cells of the central nervous system. DNA demethylation and histone modifications are two types of epigenetic modifications working in a coordinate or synergistic manner to regulate the expression of various genes. This study investigated and elucidated the epigenetic regulation of Nestin gene expression during embryonic differentiation along the neural cell lineage. Nestin exhibits differential DNA methylation and histone acetylation patterns in Nestin-expressing and nonexpressing cells. In P19 embryonic carcinoma cells, activation of Nestin expression is mediated by both trichostatin A and 5-aza-20-deoxycytidine treatment, concomitant with histone acetylation, but not with DNA demethylation. Nestin transcription is also mediated by treatment with retinoic acid, again in the absence of DNA demethylation. Thus, histone acetylation is sufficient to mediate the activation of Nestin transcription. This study proposed that the regulation of Nestin gene expression can be used as a model to study the epigenetic regulation of gene expression mediated by histone acetylation, but not by DNA demethylation. [ABSTRACT FROM AUTHOR]

Reactivation of Oct4 gene expression occurs within 2 days of fusion of somatic cells with pluripotent stem cells and within 9 days of postinfection of four transcription factors. We sought to determine whether somatic genome reprogramming is completed by the onset of Oct4 reactivation. The complex regulation of the reactivation of inactive X chromosome (Xi) serves as a model for studying reprogramming of chromatin domains. A time-course analysis of the DNA methylation, gene expression, and X inactivation-specific transcript (Xist)/Tsix RNA fluorescence in situ hybridization revealed that expression of pluripotency- and tissue-specific marker genes was reset to the level of pluripotent stem cells within 2 days of fusion, whereas reprogramming of Xist/reactivation of Xi took at least 9 days. We found that trichostatin A, which normally activates gene expression, results in downregulation of Xist. This is due to activation of Dnmt3a and Tsix, two negative regulators of Xist. Moreover, delayed reprogramming of Xist/reactivation of inactive X chromosome after cell fusion was accelerated by DNA methylation and histone deacetylation of Xist, which follow upregulation of Dnmt3a and Tsix. [ABSTRACT FROM AUTHOR]

Reprogramming of somatic cells is a valuable tool to understand the mechanisms of regaining pluripotency and further opens up the possibility of generating patient-specific pluripotent stem cells. Reprogramming of mouse and human somatic cells into pluripotent stem cells, designated as induced pluripotent stem (iPS) cells, has been possible with the expression of the transcription factor quartet Oct4 (also known as Pou5f1), Sox2, c-Myc and Klf4 (refs 1–11). Considering that ectopic expression of c-Myc causes tumorigenicity in offspring and that retroviruses themselves can cause insertional mutagenesis, the generation of iPS cells with a minimal number of factors may hasten the clinical application of this approach. Here we show that adult mouse neural stem cells express higher endogenous levels of Sox2 and c-Myc than embryonic stem cells, and that exogenous Oct4 together with either Klf4 or c-Myc is sufficient to generate iPS cells from neural stem cells. These two-factor iPS cells are similar to embryonic stem cells at the molecular level, contribute to development of the germ line, and form chimaeras. We propose that, in inducing pluripotency, the number of reprogramming factors can be reduced when using somatic cells that endogenously express appropriate levels of complementing factors. [ABSTRACT FROM AUTHOR]

The fusion of pluripotent embryonic cells with somatic cells results in reprogramming of the somatic cell genome. Oct4-green fluorescent protein (GFP) transgenes that do not contain the proximal enhancer (PE) region are widely used to visualize reprogramming of the somatic to the pluripotent cell state. The temporal onset of Oct4-GFP activation has been found to occur 40-48 hours postfusion. We asked whether activation of the transgene actually reflects activation of the endogenous Oct4 gene. In the current study, we show that activation of an Oct4-GFP transgene that contains the PE region occurs within 22 hours of fusion. In addition, demethylation of the Oct4-GFP transgene and that of the endogenous Oct4 and Nanog genes was found to occur within 24 hours of fusion. As this timing corresponds with the timing of cell cycle completion in embryonic stem cells and fusion hybrids (~22 hours), we postulate that pluripotential reprogramming of the somatic cell genome begins during the first cell cycle after the fusion of a somatic cell with a pluripotent cell and has been completed by day 2 postfusion. [ABSTRACT FROM AUTHOR]

Pluripotent cells have been suggested as a prime source to reprogram somatic cells. We used F9 EC cells as a pluripotent partner to reprogram neurosphere cells (NSCs) because they exhibit a nonneural differentiation potential in the presence of retinoic acid. F9-NSC hybrid cells displayed various features of reprogramming, such as reactivation of pluripotency genes, inactivation of tissue-specific genes, and reactivation of the inactive X chromosome. As the hybrid cells undergo differentiation, the pluripotency markers Oct4 and Nanog were downregulated. Whereas neural marker genes were not upregulated, endodermal and mesodermal markers were, suggesting that NSCs lose memory of their neural origin and preferentially differentiate to the lineages corresponding to the F9 program. After fusion, the methylation status in the Xist region was similar to that of F9 EC cells. However, upon differentiation, the Xist region failed to resume the methylation patterns of differentiated cells, suggesting that the Xist in F9-NSC hybrids does not easily acquire a differentiated state. [ABSTRACT FROM AUTHOR]

It has been established that certain heavy metals including silver have antibacterial properties, and that they may be useful components of antibacterial materials. This knowledge of the antibacterial activity of silver has already been put to use to decrease the occurrence of urinary catheter infections and to develop topical antibacterial agents for burn victims. In this study, the ability of silver-coated ceramic beads (Ag-CBs), sintered at 375, 400, 425, or 450 °C, to sterilize Sphingomonas sp. in commercial mineral water was evaluated. Over 3% (w/v) Ag-CBs in mineral water completely sterilized Sphingomonas sp. after 6 h, regardless of the sintering temperature. The Ag-CBs sintered at 400 °C were the most useful since they did not produce clouding in the water and the bacteria were effectively killed. The antimicrobial activity of the 400 °C-sintered Ag-CBs was comparable to that of free silver ions at 15 μg/l, showing that the silver ions released from the Ag-CBs resulted in the complete sterilization of the microbial cells in mineral water. These results suggest that Ag-CBs present an easy-to-use option for mineral water sterilization. [ABSTRACT FROM AUTHOR]

Green tea polyphenol (GTP) together with dimethylsulphoxide (DMSO) were added to a freezing solution of osteoblastic cells (rat calvarial osteoblasts and human osteosarcoma cells) exposed to repeated freeze/thaw cycles (FTC) to induce oxidative stress. When cells were subjected to 3 FTCs, freezing medium containing 10% (v/v) DMSO and 500 μg GTP ml−1 significantly ( p < 0.05) suppressed cell detachment and growth inhibition by over 63% and protected cell morphology. Furthermore, the alkaline phosphatase activity of osteoblastic cells was appreciably maintained after 2 and 3 FTCs in this mixture. Polyphenols may thus be of use as a cell cryopreservant and be advantageous in such fields as cell transplantation and tissue engineering. [ABSTRACT FROM AUTHOR]

Neural cortical cells, isolated from prenatal rat cerebra, were grown on surface-modified poly(lactic-co-glycolic acid, 65:35) (PLGA) films coated with poly-D-lysine (PDL) with either laminin (LN), fibronectin (FN) or collagen (CN). Immunocytochemistry showed that the isolated cells were highly immunopositive for both neurofilament and MAP-2 with well-organized neurites and somatodendritic localization. The presence of PDL with LN or FN on the PLGA films was essential for increased neural cell growth. Also, PLGA films coated with either PDL/LN or PDL/FN mixtures had higher neurite outgrowth and regular differentiation. [ABSTRACT FROM AUTHOR]

Nanowires can serve as three-dimensional platforms at the nanometer scale for highly efficient chemical energy storage and conversion vehicles, such as fuel cells and secondary batteries. Here we report a coin-type Si nanowire (NW) half-cell Li-ion battery showing the Li capacity of approximately 4000 mAh/g, which nearly approaches the theoretical limit of 4200 mAh/g, with very high Coulombic efficiency of up to 98%. Concomitantly, we provide direct evidence of reversible phase transitions in the Si NW anodes at the full electrochemical cycles, varying from pure Si to Li22Si5 phase, which has been known empirically inaccessible in the bulk limit. [ABSTRACT FROM AUTHOR]

The fusion of somatic cells with pluripotent cells results in the generation of pluripotent hybrid cells. Because the `memory' of somatic cells seems to be erased during fusion-induced reprogramming, genetic reprogramming is thought to be a largely unidirectional process. Here we show that fusion-induced reprogramming, which brings about the formation of pluripotent hybrids, does not always follow a unidirectional route. Xist is a unique gene in that it is reprogrammed to the state of somatic cells in fusion-induced pluripotent hybrids. In hybrids formed from the cell fusion of embryonal carcinoma cells (ECCs) with male neural stem cells (mNSCs), the Xist gene was found to be reprogrammed to the somatic cell state, whereas the pluripotency-related and tissue-specific marker genes were reprogrammed to the pluripotent cell state. Specifically, Xist is not expressed in hybrids, because the `memory' of the somatic cell has been retained (i.e. mNSCs do not exhibit Xist expression) and that of the pluripotent cell erased (i.e. inactivation of the partially active Xist gene of ECCs, complete methylation of the Xist region). The latter phenomenon is induced by male, but not by female, NSCs. [ABSTRACT FROM AUTHOR]

Studies examining somatic cell nuclear transfer (SCNT) have demonstrated that the cytoplasm of an oocyte can reprogram the genome of a somatic cell to an embryonic state. Reprogramming of a somatic cell to a pluripotent state can also be achieved either by fusion with embryonic stem cells or by the ectopic expression of defined transcription factors. Pluripotent stem cells derived by this means can be differentiated in defined tissues, and serve as model systems for the study and treatment of human diseases. Using the first application of stable RNA interference in human embryonic stem cells, we showed that knockdown of Oct4 and Nanog gene expression promotes their differentiation into cells expressing trophectodermal and endodermal markers, thereby demonstrating a conserved role for these factors in the self - renewal of human embryonic stem cells (Zaehres et al., 2005). We have developed retro / lentivirus vectors for efficient transgene expression in mouse and human embryonic stem cells as well as progenies derived thereof (Zaehres and Daley, 2006). We are further developing transgenic reporter systems expressing green fluorescent protein (GFP) and other selection markers under the control of mouse and human Oct4 and Nanog promoter elements. Currently we are applying these vector systems to explore reprogramming induced by fusion of somatic cells with mouse and human embryonic stem cells and to explore reprogramming of mouse and human somatic cells with defined transcription factors. These technologies are instrumental in providing a detailed, molecular understanding of the genomic reprogramming process with the ultimate goal to induce pluripotent cells in vitro and in vivo by the use of defined transcription factors and small molecules. [ABSTRACT FROM AUTHOR]

Embryonic stem cells lose pluripotency through a series of fate commitment events. We have isolated embryonic stem-derived cells that have gone through the earliest steps in neural commitment. GCNF is necessary for this commitment. Microarray profiling suggests both novel pluripotency-associated transcription factors and strategies to reprogram these cells back to pluripotency. [ABSTRACT FROM AUTHOR]

Background & aims Mammalian oocytes can be induced to cleave if extrusion of the first polar body is inhibited or reverted. The process, named parthenogenetic activation, can give rise to blastocysts. The inner cell mass of blastocysts can be isolated and used to derive parthenogenetic embryonic stem cell lines (pES). In our present study we investigated whether pES by cell fusion can reprogram somatic cells to acquire a pluripotent state. Methods & results As a somatic fusion partner we chose neural stem cells (NSC) that carry two transgenes, Oct4-GFP and Rosa26-Neo. After fusion of these cells with pES cells, colonies appeared that were GFP-positive and neomycin-resistant. The presence of such colonies showed reactivation of Oct4 expression, suggesting reprogramming of NSCs. The fused cells were further investigated by RT-PCR and immunofluorescence for the presence of Oct4 and Nanog. Ploidy analyzes confirmed the tetraploid state of the pES-NSC. After transplantation into SCID-mice pES as well as pES/NSC gave rise to teratomas, comprising cells from all three germ layers. In addition, several tests, such as DNA-methylation, were performed to demonstrate that reprogramming in the pES-NSC fusion hybrids had occurred. Conclusion The pES has the reprogramming potential to convert somatic cells into a pluripotent state. Therefore, pES might be an alternative source for reprogramming strategies such as fusion with somatic cells as well as for the isolation of reprogramming factors from cell extracts. [ABSTRACT FROM AUTHOR]