Distinct characteristics of two types of alternative lengthening of telomeres in mouse embryonic stem cells.

Telomere length must be maintained in actively dividing cells to avoid cellular arrest or death. In the absence of telomerase activity, activation of alternative lengthening of telomeres (ALT) allows the maintenance of telomeric length and prolongs the cellular lifespan. Our previous studies have established two types of ALT survivors from mouse embryonic stem cells. The key differences between these ALT survivors are telomere-constituting sequences: non-telomeric sequences and canonical telomeric repeats, with each type of ALT survivors being referred to as type I and type II, respectively. We explored how the characteristics of the two types of ALT lines reflect their fates using multi-omics approaches. The most notable gene expression signatures of type I and type II ALT cell lines were chromatin remodelling and DNA repair, respectively. Compared with type II cells, type I ALT cells accumulated more mutations and demonstrated persistent telomere instability. These findings indicate that cells of the same origin have separate routes for survival, thus providing insights into the plasticity of crisis-suffering cells and cancers.

Telomeres reforged with non-telomeric sequences in mouse embryonic stem cells.

Telomeres are part of a highly refined system for maintaining the stability of linear chromosomes. Most telomeres rely on simple repetitive sequences and telomerase enzymes to protect chromosomal ends; however, in some species or telomerase-defective situations, an alternative lengthening of telomeres (ALT) mechanism is used. ALT mainly utilises recombination-based replication mechanisms and the constituents of ALT-based telomeres vary depending on models. Here we show that mouse telomeres can exploit non-telomeric, unique sequences in addition to telomeric repeats. We establish that a specific subtelomeric element, the mouse template for ALT (mTALT), is used for repairing telomeric DNA damage as well as for composing portions of telomeres in ALT-dependent mouse embryonic stem cells. Epigenomic and proteomic analyses before and after ALT activation reveal a high level of non-coding mTALT transcripts despite the heterochromatic nature of mTALT-based telomeres. After ALT activation, the increased HMGN1, a non-histone chromosomal protein, contributes to the maintenance of telomere stability by regulating telomeric transcription. These findings provide a molecular basis to study the evolution of new structures in telomeres.

Repair and Reconstruction of Telomeric and Subtelomeric Regions and Genesis of New Telomeres: Implications for Chromosome Evolution.

DNA damage repair within telomeres are suppressed to maintain the integrity of linear chromosomes, but the accidental activation of repairs can lead to genome instability. This review develops the concept that mechanisms to repair DNA damage in telomeres contribute to genetic variability and karyotype evolution, rather than catastrophe. Spontaneous breaks in telomeres can be repaired by telomerase, but in some cases DNA repair pathways are activated, and can cause chromosomal rearrangements or fusions. The resultant changes can also affect subtelomeric regions that are adjacent to telomeres. Subtelomeres are actively involved in such chromosomal changes, and are therefore the most variable regions in the genome. The case of Caenorhabditis elegans in the context of changes of subtelomeric structures revealed by long-read sequencing is also discussed. Theoretical and methodological issues covered in this review will help to explore the mechanism of chromosome evolution by reconstruction of chromosomal ends in nature.

Enhancement of Glucose Uptake by Meso-Dihydroguaiaretic Acid through GLUT4 Up-Regulation in 3T3-L1 Adipocytes.

Type 2 diabetes is characterized by insulin resistance, which leads to increased blood glucose levels. Adipocytes are involved in the development of insulin resistance, resulting from the dysfunction of the insulin signaling pathway. In this study, we investigated whether meso-dihydroguaiaretic acid (MDGA) may modulate glucose uptake in adipocytes, and examined its mechanism of action. MDGA enhanced adipogenesis through up-regulation of peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α in 3T3-L1 adipocytes partially differentiated with sub-optimal concentrations of insulin. MDGA also increased glucose uptake by stimulating expression and translocation of glucose transporter 4 (GLUT4) in adipocytes. These results suggest that MDGA may increase GLUT4 expression and its translocation by promoting insulin sensitivity, leading to enhanced glucose uptake.

Internal genomic regions mobilized for telomere maintenance in C. elegans.

Because DNA polymerase cannot replicate telomeric DNA at linear chromosomal ends, eukaryotes have developed specific telomere maintenance mechanisms (TMMs). A major TMM involves specialized reverse transcriptase, telomerase. However, there also exist various telomerase-independent TMMs (TI-TMMs), which can arise both in pathological conditions (such as cancers) and during evolution. The TI-TMM in cancer cells is called alternative lengthening of telomeres (ALT), whose mechanism is not fully understood. We generated stably maintained telomerase-independent survivors from C. elegans telomerase mutants and found that, unlike previously described survivors in worms, these survivors "mobilize" specific internal sequence blocks for telomere lengthening, which we named TALTs (templates for ALT). The cis-duplication of internal genomic TALTs produces "reservoirs" of TALTs, whose trans-duplication occurs at all chromosome ends in the ALT survivors. Our discovery that different TALTs are utilized in different wild isolates provides insight into the molecular events leading to telomere evolution.

Telomere maintenance through recruitment of internal genomic regions.

Cells surviving crisis are often tumorigenic and their telomeres are commonly maintained through the reactivation of telomerase. However, surviving cells occasionally activate a recombination-based mechanism called alternative lengthening of telomeres (ALT). Here we establish stably maintained survivors in telomerase-deleted Caenorhabditis elegans that escape from sterility by activating ALT. ALT survivors trans-duplicate an internal genomic region, which is already cis-duplicated to chromosome ends, across the telomeres of all chromosomes. These 'Template for ALT' (TALT) regions consist of a block of genomic DNA flanked by telomere-like sequences, and are different between two genetic background. We establish a model that an ancestral duplication of a donor TALT region to a proximal telomere region forms a genomic reservoir ready to be incorporated into telomeres on ALT activation.

Isolated central nervous system relapse of acute lymphoblastic leukemia.

Acute lymphoblastic leukemia (ALL) is the most common form of childhood cancer and may exhibit central nervous system (CNS) involvement. Advances in chemotherapy and effective CNS prophylaxis have significantly decreased the incidence of CNS relapse of ALL to 5-10%. Here, we report the case of a patient with isolated CNS relapse of standard risk group pre-B-cell type ALL in an 11-year-old girl, relapsed 3 years after successful completion of chemotherapy. An 11-year-old girl visited our hospital complaining of headache, dizziness, vomiting, and visual field defects. Neurological examination revealed left-side homonymous hemianopsia. Brain magnetic resonance imaging showed a large irregular dural-based sulcal hematoma in the right parietal and occipital lobes. Surgery to remove the hematoma revealed the existence of hematopoietic malignancy after pathologic evaluation. Bone marrow biopsy was subsequently performed but showed no evidence of malignancy.

Clinical significance of radiological stability in reconstructed thoracic and lumbar spine following vertebral body resection.

OBJECTIVE: Vertebral body replacement following corpectomy in thoracic or lumbar spine is performed with titanium mesh cage (TMC) containing any grafts. Radiological changes often occur on follow-up. This study investigated the relationship between the radiological stability and clinical symptoms. METHODS: The subjects of this study were 28 patients who underwent corpectomy on the thoracic or lumbar spine. Their medical records and radiological data were retrospectively analyzed. There were 23 cases of tumor, 2 cases of trauma, and 3 cases of infection. During operation, spinal reconstruction was done with TMC and additional screw fixation. We measured TMC settlement in sagittal plane and spinal angular change in coronal and sagittal plane at postoperative one month and last follow-up. Pain score was also checked. We investigated the correlation between radiologic change and pain status. Whether factors, such as the kind of graft material, surgical approach, and fusion can affect the radiological stability or not was analyzed as well. RESULTS: Mean follow-up was 23.6 months. During follow-up, 2.08±1.65° and 6.96±2.08° of angular change was observed in coronal and sagittal plane, respectively. A mean of cage settlement was 4.02±2.83 mm. Pain aggravation was observed in 4 cases. However, no significant relationship was found between spinal angular change and pain status (p=0.518, 0.458). Cage settlement was seen not to be related with pain status, either (p=0.644). No factors were found to affect the radiological stability. CONCLUSION: TMC settlement and spinal angular change were often observed in reconstructed spine. However, these changes did not always cause postoperative axial pain.

Evaluation of risk factors for vertebral compression fracture after stereotactic radiosurgery in spinal tumor patients.

OBJECTIVE: Stereotactic radiosurgery (SRS) is an emerging treatment modality for malignant spinal tumors. After SRS, some patients suffered from pain aggravation due to development of vertebral compression fracture (VCF). In these cases, surgery should be considered. METHODS: This study consisted of 72 patients who underwent SRS due to spinal tumors. In them, whether post-SRS VCF developed or not was investigated. We retrospectively analyzed their medical records and radiological imaging data. VCF was diagnosed with X-ray and magnetic resonance imaging (MRI). The incidence, time to development and risk factors for VCF were investigated. Age, sex, whole vertebral body involvement rate, vertebral body osteolysis rate, pre-SRS spinal deformity, spinal instability neoplastic score (SINS), spinal canal encroachment, lesion level, and radiation dose were analyzed as potential risk factors. A multi-variate logistic regression model was used for statistical analysis. RESULTS: In our study population, VCF was observed in 26 patients (36%). The mean time to VCF development was 1.5 months. Using uni-variate analyses, the significant risk factors were pre-SRS spinal deformity, SINS, vertebral body osteolysis rate, and whole vertebral body involvement rate. However, using multi-variate analyses, the only significant risk factor was vertebral body osteolysis rate. The patients whose vertebral body was destroyed by more than 60% showed an 8.4 times higher risk of VCF than those who had vertebral body destruction of less than 60%(p=0.016). CONCLUSION: The most significant prognostic factor for post-SRS VCF was vertebral body osteolysis rate, rather than whole vertebral body involvement rate. When more than 60% of the vertebral body was destroyed, the risk of VCF or spinal deformity was high.

Neuroprotective benzyl benzoate glycosides from Disporum viridescens roots in HT22 hippocampal neuronal cells.

Bioassay-guided fractionation of the EtOAc extract from Disporum viridescens roots led to the isolation of five new benzyl benzoate glycosides, BBGs (1-5). The neuroprotective activities of the BBGs were screened using neuronal HT22 hippocampal cells. BBG-D (4) significantly protected murine hippocampal HT22 cells against glutamate-induced neurotoxicity by maintaining the antioxidative defense systems such as superoxide dismutase, glutathione reductase, glutathione peroxidase, and the glutathione content. BBG-D, in a dose-and time-dependent manner, increased HO-1 expression through the selective activation of pERK signaling among the MAPK pathways. These results suggest that BBG-D could be a promising candidate for the treatment of neurodegenerative diseases related to glutamate-induced oxidative neuronal cytotoxicity.

Simvastatin suppresses RANTES-mediated neutrophilia in polyinosinic-polycytidylic acid-induced pneumonia.

Recently, statins have been shown to have anti-inflammatory effects on lung inflammatory diseases. However, the mechanisms of action of simvastatin in viral pneumonia have yet to be elucidated, although viral infection remains a considerable health threat. In this study, we hypothesised that simvastatin inhibits polyinosinic-polycytidylic acid (poly I:C)-induced airway inflammation, such as RANTES (regulated on activation, normal T-cell expressed and secreted) expression and inflammatory cell recruitment. In bronchial cells, the effect of simvastatin on poly I:C-induced RANTES expression and signal transducer and activator of transcription (STAT)3-mediated signal transduction was determined using an ELISA and short hairpin (sh)RNA system. In a poly I:C-induced pneumonia mouse model, immunological changes in the lungs after simvastatin inhalation, such as inflammatory cell recruitment and cytokine/chemokine release, were examined. In poly I:C-stimulated bronchial cells, RANTES secretion was increased by STAT3 activation, and simvastatin suppressed poly I:C-induced STAT3 activation, resulting in inhibition of RANTES expression. In BALB/c mice stimulated with inhaled poly I:C, RANTES expression and neutrophil infiltration into the airway were elevated. However, simvastatin treatment attenuated STAT3 activation, RANTES release and subsequent neutrophilia in the lungs. These findings suggest that simvastatin inhibits airway inflammation, but there are other mechanisms that need to be fully elucidated.

Anti-dermatitis effects of oak wood vinegar on the DNCB-induced contact hypersensitivity via STAT3 suppression.

AIMS OF THE STUDY: The aim of this study was to evaluate the anti-dermatitis effects of oak wood vinegar (OWV) in a 2,4-dinitrochlorobenzene (DNCB)-induced contact dermatitis mice model. MATERIALS AND METHODS: Immunoglobulin E (IgE) production, infiltration of immune cells (neutrophils, CD3+ cells), inducible nitric oxide synthase (iNOS) expression, skin thickness, and expression of phosphorylated STAT3 (signal transducers and activators of transcription 3) protein were tested in a DNCB-induced contact dermatitis model. In vitro wound healing and proliferative assays were also performed. RESULTS: OWV showed anti-inflammatory effects on DNCB-induced dermatitis in mice, leading to inhibition of IgE production, immune cell infiltration, and iNOS expression. Skin thickness and the level of phospho-STAT3 were dramatically reduced by OWV. Using the HaCaT human keratinocyte cell line, we confirmed that constitutive STAT3 activation induced faster proliferation of epithelial cells. In addition, OWV suppressed HaCaT proliferative ability and phospho-STAT3 levels. CONCLUSIONS: The study revealed that OWV has anti-inflammatory and anti-proliferative effects in a DNCB-induced contact dermatitis mice model. Furthermore, we showed that the mechanism by which OWV most likely inhibits epithelial proliferation is through STAT3 inactivation.

Magnolia officinalis reverses alcoholic fatty liver by inhibiting the maturation of sterol regulatory element-binding protein-1c.

The generally accepted hypothesis for the pathogenesis of alcoholic liver disease (ALD) is the two-hit model, which proposes that fat accumulation in the liver increases the sensitivity of the liver to a second hit that leads to inflammatory liver cell damage. In this study we evaluated the effects of Magnolia officinalis (MO), which contains honokiol and magnolol as the primary pharmacological components, to eradicate fatty liver in rats fed an ethanol diet. In vitro studies showed that MO was able to protect RAW 264.7 cells from ethanol-induced production of tumor necrosis factor-alpha, reactive oxygen species, and superoxide anion radicals; the activation of NADPH oxidase; and subsequent cell death. We also investigated the therapeutic effects of MO on alcoholic fatty liver in Lieber-DeCarli ethanol diet-fed rats. MO treatment of the rats for the last 2 weeks of ethanol feeding completely reversed all the serum, hepatic parameters, and fatty liver changes. The increased maturation of sterol regulatory element-binding protein-1c in the liver by ethanol treatment was completely inhibited by treatment with MO. Therefore, MO may be a promising candidate for development as a therapeutic agent for ALD.