Enhanced Intervertebral Disc Repair via Genetically Engineered Mesenchymal Stem Cells with Tetracycline Regulatory System.

The therapeutic potential of Mesenchymal stem cells (MSCs) for the treatment of Intervertebral disc (IVD) degeneration can be enhanced by amplifying specific cytokines and proteins. This study aimed to investigate the therapeutic potential of tetracycline-off system-engineered tonsil-derived mesenchymal stem cells (ToMSC-Tetoff-TGFß1-IGF1-BMP7) for treating intervertebral disc (IVD) degeneration. ToMSCs were isolated from a tonsillectomy patient and genetically modified with four distinct plasmids via CRISPR/Cas9-mediated knock-in gene editing. Transgene expression was confirmed through immunofluorescence, western blots, and an enzyme-linked immunosorbent assay for transforming growth factor beta 1 (TGFß1) protein secretion, and the effect of MSC-TetOff-TGFß1-IGF1-BMP7 on disc injury was assessed in a rat model. The ToMSC-Tetoff-TGFß1-IGF1-BMP7 treatment exhibited superior therapeutic effects compared to ToMSC-TGFß1, and ToMSC-SDF1α implantation groups, stimulating the regeneration of nucleus pulposus (NP) cells crucial for IVD. The treatment showed potential to restore the structural integrity of the extracellular matrix (ECM) by upregulating key molecules such as aggrecan and type II collagen. It also exhibited anti-inflammatory properties and reduced pain-inducing neuropeptides. ToMSC-Tetoff-TGFß1-IGF1-BMP7 holds promise as a novel treatment for IVD degeneration. It appears to promote NP cell regeneration, restore ECM structure, suppress inflammation, and reduce pain. However, more research and clinical trials are required to confirm its therapeutic potential.

Tumor Necrosis Factor-producing T-regulatory Cells Are Associated With Severe Liver Injury in Patients With Acute Hepatitis A.

BACKGROUND AND AIMS: CD4+CD25+Foxp3+ T-regulatory (Treg) cells control immune responses and maintain immune homeostasis. However, under inflammatory conditions, Treg cells produce cytokines that promote inflammation. We investigated production of tumor necrosis factor (TNF) by Treg cells in patients with acute hepatitis A (AHA), and examined the characteristics of these cells and association with clinical factors. METHODS: We analyzed blood samples collected from 63 patients with AHA at the time of hospitalization (and some at later time points) and 19 healthy donors in South Korea. Liver tissues were collected from patients with fulminant AHA during liver transplantation. Peripheral blood mononuclear cells were isolated from whole blood and lymphocytes were isolated from liver tissues and analyzed by flow cytometry. Cytokine production from Treg cells (CD4+CD25+Foxp3+) was measured by immunofluorescence levels following stimulation with anti-CD3 and anti-CD28. Epigenetic stability of Treg cells was determined based on DNA methylation patterns. Phenotypes of Treg cells were analyzed by flow cytometry and an RORγt inhibitor, ML-209, was used to inhibit TNF production. Treg cell suppression assay was performed by co-culture of Treg-depleted peripheral blood mononuclear cells s and isolated Treg cells. RESULTS: A higher proportion of CD4+CD25+Foxp3+ Treg cells from patients with AHA compared with controls produced TNF upon stimulation with anti-CD3 and anti-CD28 (11.2% vs 2.8%). DNA methylation analysis confirmed the identity of the Treg cells. TNF-producing Treg cells had features of T-helper 17 cells, including up-regulation of RORγt, which was required for TNF production. The Treg cells had reduced suppressive functions compared with Treg cells from controls. The frequency of TNF-producing Treg cells in AHA patients' blood correlated with their serum level of alanine aminotransferase. CONCLUSIONS: Treg cells from patients with AHA have altered functions compared with Treg cells from healthy individuals. Treg cells from patients with AHA produce higher levels of TNF, gain features of T-helper 17 cells, and have reduced suppressive activity. The presence of these cells is associated with severe liver injury in patients with AHA.

Dynamic Long-Range Chromatin Interaction Controls Expression of IL-21 in CD4+ T Cells.

IL-21, a pleiotropic cytokine strongly linked with autoimmunity and inflammation, regulates diverse immune responses. IL-21 can be potently induced in CD4(+) T cells by IL-6; however, very little is known about the mechanisms underlying the transcriptional regulation of the Il21 gene at the chromatin level. In this study, we demonstrated that a conserved noncoding sequence located 49 kb upstream of the Il21 gene contains an enhancer element that can upregulate Il21 gene expression in a STAT3- and NFAT-dependent manner. Additionally, we identified enhancer-blocking insulator elements in the Il21 locus, which constitutively bind CTCF and cohesin. In naive CD4(+) T cells, these upstream and downstream CTCF binding sites interact with each other to make a DNA loop; however, the Il21 promoter does not interact with any cis-elements in the Il21 locus. In contrast, stimulation of CD4(+) T cells with IL-6 leads to recruitment of STAT3 to the promoter and novel distal enhancer region. This induces dynamic changes in chromatin configuration, bringing the promoter and the regulatory elements in close spatial proximity. The long-range interaction between the promoter and distal enhancer region was dependent on IL-6/STAT3 signaling pathway but was disrupted in regulatory T cells, where IL-21 expression was repressed. Thus, our work uncovers a novel topological chromatin framework underlying proper transcriptional regulation of the Il21 gene.

THICKNESS OF THE MACULA, RETINAL NERVE FIBER LAYER, AND GANGLION CELL-INNER PLEXIFORM LAYER IN THE AGE-RELATED MACULAR DEGENERATION: The Repeatability Study of Spectral Domain Optical Coherence Tomography.

PURPOSE: To determine the repeatability of measuring the thickness of the central macula, retinal nerve fiber layer, and ganglion cell-inner plexiform layer (GC-IPL) using spectral domain optical coherence tomography (Cirrus HD-OCT) in eyes with age-related macular degeneration. METHODS: One hundred and thirty-four eyes were included. The measurement repeatability was assessed by an experienced examiner who performed two consecutive measurements using a 512 × 128 macular cube scan and a 200 × 200 optic disk cube scan. To assess changes in macular morphology in patients with age-related macular degeneration, the patients were divided into the following three groups according to the central macular thickness (CMT): A group, CMT < 200 µm; B group, 200 µm ≤ CMT < 300 µm; and C group, CMT > 300 µm. RESULTS: Measurement repeatability was assessed using test-retest variability, a coefficient of variation, and an intraclass correlation coefficient. The mean measurement repeatability for the central macular, retinal nerve fiber layer, and GC-IPL thickness was high in the B group. The mean measurement repeatability for both the central macula and retinal nerve fiber layer thickness was high in the A and C groups, but was lower for the GC-IPL thickness. The measurement repeatability for GC-IPL thickness was high in the B group, but low in the A group and in the C group. CONCLUSION: The automated measurement repeatability for GC-IPL thickness was significantly lower in patients with age-related macular degeneration with out of normal CMT range. The effect of changes in macular morphology should be considered when analyzing GC-IPL thicknesses in a variety of ocular diseases.

Effect of Iris registration on visual outcome in wavefront-guided LASEK for myopic astigmatism.

PURPOSE: This study aimed to investigate the effect of iris registration (IR) on visual outcomes in wavefront-guided LASEK for myopic astigmatism. METHODS: The retrospective chart review was performed for wavefront-guided LASEK using VISX Star S4 in patients with myopic astigmatism (cylinder ≥ 1.00 diopter[D]). Eyes were divided into IR group (LASEK with IR at the time of surgery) and Non-IR group (LASEK without IR system + failed-IR engagement during LASEK). Visual acuity (VA), astigmatism, higher-order aberration (HOA), and contrast sensitivity were assessed preoperatively and 3 months postoperatively. The IR and Non-IR groups were subcategorized depending on the spherical equivalent (lower myopia ≤-5.00 D vs. higher myopia >-5.00 D) for the comparison of HOA changes. RESULTS: Postoperative uncorrected VAs showed no differences between IR (n = 30) and Non-IR (n = 46). In astigmatic vector analyses, no differences were noted in the mean magnitude of error and the mean angle of error between two groups. There were no differences in postoperative total HOA, spherical aberration (SA), coma, and trefoil between the groups, either. The total HOA and SA increased in both groups, while coma increased only in Non-IR. In higher myopia, ΔRMS of coma was smaller in IR. Preoperative and postoperative total HOA were linearly correlated in Non-IR, but not for IR. Contrast sensitivity of 12 cycles per degree improved in both groups. CONCLUSION: IR had similar outcomes to conventional trackers in wavefront-guided LASEK, with less tendency of inducing coma, especially in higher myopia.

Prospective, Multicenter Study on Tinnitus Changes after Cochlear Implantation.

OBJECTIVE: To investigate the time course of tinnitus changes in patients receiving cochlear implantation (CI) in a prospective, multicenter setting and to determine related factors. MATERIALS AND METHODS: A total of 79 adult patients who underwent CI were included in this study. We used the same questionnaires sequentially 5 times. The questionnaires included the Visual Analog Scale (VAS) for tinnitus severity, the Tinnitus Handicap Inventory (THI), Beck's Depression Index (BDI), and the Brief Encounter Psychosocial Instrument (BEPSI) for stress assessment. RESULTS: Tinnitus was present in 59 (74.7%) of the 79 study subjects. After CI, tinnitus was eliminated in 10 patients (25%) and improved in 16 patients (40%) of the 40 patients who completed the final questionnaires, and most of the tinnitus reduction occurred in the early period of CI use. In an analysis of psychological functioning with CI, BDI was reduced significantly after CI. Multiple linear regression analysis revealed that preoperative auditory steady-state response (ASSR), THI, and final BDI score were significantly associated with the changes in tinnitus after CI. CONCLUSIONS: Most of the tinnitus reduction occurred within 1 month after CI use, and the changes were significantly associated with THI, ASSR, and BDI scores 6 months after CI. CI is a valuable therapeutic modality in tinnitus of a deafened ear.

Staufen1-mediated mRNA decay induces Requiem mRNA decay through binding of Staufen1 to the Requiem 3'UTR.

Requiem (REQ/DPF2) was originally identified as an apoptosis-inducing protein in mouse myeloid cells and belongs to the novel Krüppel-type zinc finger d4-protein family of proteins, which includes neuro-d4 (DPF1) and cer-d4 (DPF3). Interestingly, when a portion of the REQ messenger ribonucleic acid (mRNA) 3' untranslated region (3'UTR), referred to as G8, was overexpressed in K562 cells, ß-globin expression was induced, suggesting that the 3'UTR of REQ mRNA plays a physiological role. Here, we present evidence that the REQ mRNA 3'UTR, along with its trans-acting factor, Staufen1 (STAU1), is able to reduce the level of REQ mRNA via STAU1-mediated mRNA decay (SMD). By screening a complementary deoxyribonucleic acid (cDNA) expression library with an RNA-ligand binding assay, we identified STAU1 as an interactor of the REQ mRNA 3'UTR. Specifically, we provide evidence that STAU1 binds to putative 30-nucleotide stem-loop-structured RNA sequences within the G8 region, which we term the protein binding site core; this binding triggers the degradation of REQ mRNA and thus regulates translation. Furthermore, we demonstrate that siRNA-mediated silencing of either STAU1 or UPF1 increases the abundance of cellular REQ mRNA and, consequently, the REQ protein, indicating that REQ mRNA is a target of SMD.

CCCTC-binding factor controls the homeostatic maintenance and migration of Langerhans cells.

BACKGROUND: Langerhans cells (LCs) are skin-resident dendritic cells (DCs) that orchestrate skin immunity. CCCTC-binding factor (CTCF) is a highly conserved DNA-binding protein that regulates higher-order chromatin organization and is involved in various gene regulation processes. OBJECTIVE: We sought to clarify a possible role for CTCF in LC homeostasis and function in vivo. METHODS: We used a conditional gene deletion mouse system to generate DC- and LC-specific CTCF-ablated mice. Short hairpin RNA-mediated RNA interference was used to silence CTCF expression in human monocyte-derived Langerhans cells. DC populations were assessed by using flow cytometry and immunofluorescence. Gene expression arrays were performed to identify genes regulated by CTCF in LCs. Contact hypersensitivity and epicutaneous sensitization responses were measured to examine the functional significance of CTCF ablation. RESULTS: DC-specific CTCF deletion led to a reduced pool of systemic DCs, with LCs most severely affected. Decreases in epidermal LC numbers were specifically associated with self-turnover defects. Interestingly, CTCF-deficient LCs demonstrated impaired migration out of the epidermis. Whole-transcriptome analyses revealed that genes that promoted cell adhesion were highly expressed, but CCR7 was downregulated in CTCF-depleted LCs. Hapten-induced contact hypersensitivity responses were more sustained in LC-specific CTCF-deficient mice, whereas epicutaneous sensitization to protein antigen was attenuated, indicating that CTCF-dependent LC homeostasis is required for optimal immune function of LCs in a context-dependent manner. CONCLUSION: Our results show that CTCF positively regulates the homeostatic pool and the efficient emigration of LCs, which are required for modulating the functional immune network of the skin.

Adverse effects of low-dose systemic cyclosporine therapy in high-risk penetrating keratoplasty.

PURPOSE: The purpose of this study was to investigate the adverse effects of low-dose oral cyclosporine (CsA) therapy following high-risk corneal transplantation. METHODS: The medical records from 88 subjects who had undergone high-risk penetrating keratoplasties and had been administered oral CsA were retrospectively analyzed. High risk was defined as a history of graft rejection, three or more quadrants of vascularization, or the presence or history of intraocular inflammation. An initial CsA dose of 3-5 mg/kg per day was given for 3-7 days, followed by 2.5-3.5 mg/kg per day for approximately 1 month. The concentration of CsA was maintained at the target trough level of 120-150 ng/ml for at least 6 months or until serious complications developed. The relationship between the cumulative dose and duration of CsA administration and the adverse systemic effects, including the frequency of herpes keratitis, was evaluated. The incidence of herpes keratitis in the study subjects was compared with the incidence in 185 patients who had not received CsA therapy following penetrating keratoplasty. RESULTS: The mean survival time of the grafts was 33.6 months. Adverse effects occurred in 81.8 % of subjects. Hypertension, elevated liver enzyme levels, elevated serum creatinine level, and decreased absolute neutrophil count (ANC) were observed in 14.8, 6.8, 5.7, and 5.7 % of subjects, respectively. Simvastatin-induced rhabdomyolysis also developed in one case. Some patients exhibited minor complications, with gastrointestinal problems and hypertrichosis recorded in 5.7 and 3.4 % of subjects, respectively. Hypertension and hepatotoxicity most frequently occurred after 4 to 8 weeks of medication, while ANC decrease and nephrotoxicity generally developed after 24 weeks of treatment, with incidence related to the cumulative dose. Herpes keratitis occurred more frequently (31.8 %) in the CsA-treated subjects than in subjects that did not receive CsA therapy (p = 0.005). Most of the adverse effects were reversed after discontinuation of CsA therapy. CONCLUSION: The results of this study suggest that low-dose oral CsA therapy may induce various adverse effects, the most common of which are herpes keratitis and hypertension.

The attitude toward xenocorneal transplantation in wait-listed subjects for corneal transplantation in Korea.

Shortage of donor cornea is a significant problem in Asia, and xenocorneal transplantation is being actively studied to alleviate this problem. However, the attitudes of subjects who await corneal transplants toward xenocorneal transplantation are not known at all. Thus, this study aimed to investigate the attitudes of subjects on the waiting lists for corneal transplants, toward corneal xenotransplantation. A telephone questionnaire survey comprising six items was conducted in 132 subjects among the wait-listed individuals (n = 590) who were awaiting corneal transplantation or had undergone corneal transplantation at Seoul National University Hospital from July, 2003 to August, 2012. Among six inquiries, four questions were used to analyze attitudes toward corneal xenotransplantation. Each question pertained to (1) the acceptance of xenocorneal transplantation, (2) willingness to participate in clinical trials, (3) worries in xenocorneal transplantation, and (4) the concern of self-identity or social life after xenocorneal transplantation. To analyze demographic factors influencing the question, the subjects were arbitrarily divided into two groups: the young (age < 60 yr, n = 58) and the elderly (age ≥ 60 yr, n = 74) or the less-educated (n = 53) and the well-educated with high school diploma, college graduation, or higher education (n = 79). Collected demographic data were analyzed as influencing factors on each question using a chi-square and logistic regression tests. In this study, 42.4% of the subjects (n = 56) expressed favorable views on xenocorneal transplantation using porcine corneas to cure visual loss from corneal blindness. Among those subjects expressing favorable views (n = 56), the willingness to participate in clinical trials, knowing they and their spouses must undergo long-term surveillance, was 62.5% (n = 35). There were 76.5% of subjects (n = 101) expressing worries regarding xenocorneal transplantation, while 28.8% of subjects (n = 38) expressed their concerns about self-identity or social life after xenotransplantation. Younger subjects expressed more worry about xenotransplantation than elderly subjects. The well-educated expressed less concern over self-identity and social life than the less-educated. This survey among subjects who are wait-listed for corneal transplant or who have received a corneal transplant demonstrates that there is an interest in xenocorneal transplantation as an alternate procedure, although there are worries about the procedure that should be further explored in educational campaigns and future studies of the general population.

Cross-reactivity between decellularized porcine corneal lamellae for corneal xenobridging and subsequent corneal allotransplants.

The purpose of this study is to investigate cross-reactivity between hypertonic saline-treated decellularized porcine corneal lamellae for corneal xenobridging and subsequent corneal allotransplants. Five Chinese rhesus macaques, which had undergone anterior partial thickness corneal transplantation using hypertonic saline-treated decellularized porcine corneal lamellae in preceding experiments, were used as recipients for subsequent full-thickness corneal allografts. To determine whether sensitization of recipients to xenoantigens leads to cross-reactivity against alloantigens, we compared; (i) allogeneic one-way mixed lymphocyte reaction (MLR) of peripheral blood mononuclear cells (PBMCs) from xeno-sensitized recipients with that of PBMCs from naïve rhesus macaques, and (ii) amounts of IgG antibodies that bound to the PBMCs of a rhesus panel (five monkeys) before and after xeno-sensitization. Graft survival and immunologic profiles including memory T-cell subsets and donor rhesus-specific antibodies were also evaluated. No hyperacute or acute rejection was observed within a month of subsequent allotransplantation in any recipient. Alloreactivity by MLR was not different between xeno-sensitized rhesus recipients and naïve rhesus monkeys. Panel-reactive IgG antibodies were unchanged after xeno-sensitization, and no change in donor rhesus-specific antibodies was observed in any recipient. No significant changes in memory T-cell subsets were observed during the early post-operative period in any recipient. Decellularized porcine corneal lamellae may not increase cross-reactivity to alloantigens, and thus, porcine corneal lamellae may be used as a bridge to subsequent corneal allografting.

CD39-mediated effect of human bone marrow-derived mesenchymal stem cells on the human Th17 cell function.

This study investigated the immune-modulatory effects of human bone marrow-derived mesenchymal stem cells (hBMSCs) on human Th17 cell function through the CD39-mediated adenosine-producing pathway. The suppressive effects of hBMSCs were evaluated by assessing their effects on the proliferation of Th17 cells and the secretion of interferon (IFN)-γ and interleukin (IL)-17A by Th17 cells with or without anti-CD39 treatment. Changes in CD39 and CD73 expression on the T cells with or without co-culture of hBMSCs were evaluated by flow cytometry. hBMSCs effectively suppressed the proliferation of Th17 cells and the secretion of both IL-17A and IFN-γ from Th17 cells using by both flow cytometry and ELISA, while anti-CD39 treatment significantly reduced the inhibitory effects of hBMSCs on the proliferation and secretion of the Th17 cells. The hBMSCs induced increased expression of the CD39 and CD73 on T cells correlated with the suppressive function of hBMSCs, which was accompanied by increased adenosine production. Our data suggests that hBMSCs can effectively suppress immune responses of the Th17 cells via the CD39-CD73-mediated adenosine-producing pathway.

Ethical and regulatory guidelines in clinical trials of xenocorneal transplantation in Korea; the Korean xenocorneal transplantation consensus statement.

BACKGROUND: To establish the consensus about the conditions for undertaking clinical trials in xenocorneal transplantation in Korea, specific issues regarding the xenocorneal transplantation on ethical and regulatory aspects are addressed, and the guidelines to conduct clinical trial of the xenocorneal transplantation are proposed. METHOD AND RESULTS: Chapter 1 reviews the key ethical requirements and progress of a Korean regulatory framework for clinical trials of xenocorneal transplantation. Chapters 2-7 provide recommendations on source pigs, quality control of porcine corneal procurement, preclinical efficacy required to justify a clinical trial, strategies to prevent transmission of porcine endogenous retrovirus (PERV), patient selection for clinical trials, and informed consent in xenocorneal transplantation using either cellularized or decellularized porcine graft, which are essentially based on the International Xenotransplantation Association (IXA) islet xenotransplantation consensus statement. The consensus statement of the inclusion criteria for the patients' selection has been made by the executive board members in Korean External Eye Disease Society. CONCLUSIONS: This consensus statement will be a good initiative for Korean Food and Drug Administration to discuss final regulatory guidelines in conducting clinical trials of xenocorneal transplantation in Korea and for International Xenotransplantation Association to develop International Consensus Standards of Xenocorneal Transplantation.

Significance of osteoporosis in facial bone density using computed tomography.

OBJECTIVES: The objectives of this study were to compare the variations of bone density in the midfacial bones as measured by computed tomography (CT) scans between the osteoporosis and control groups and to evaluate the regions that facial trauma and iatrogenic problem often occur in the midface. METHODS: The 96 patients who underwent both osteomeatal unit CT scans and dual-energy x-ray absorptiometry at our hospital were included in this study retrospectively. Seven skeletal regions were chosen for evaluation: group A (orbital floor, nasal bone), group B (zygomaticomaxillary suture, zygomatic arch, zygomaticofrontal suture), and group C (anterior wall of the maxillary sinus, maxillary process). Forty-seven patients were in the osteoporosis group, and 49 patients were in the control group. On a PACS (picture archiving communication system), the region of interest was analyzed, and the Hounsfield units were measured. RESULTS: There was a significant difference in the mean bone density of the midfacial bones between the osteoporosis group and the control group (P < 0.01). For both groups, each of comparison of the 7 skeletal regions was greater as group A < group B < group C in this order (P < 0.01). CONCLUSIONS: We can see the independent effects of osteoporosis on the midfacial bones using CT scans. Estimated Hounsfield unit through CT scan is able to explain osteoporosis, which may be useful in the clinical fields in the future.

Unsubtracted Roadmap Technique for Preventing Cement Leakage During Percutaneous Vertebroplasty: A Retrospective Cohort Study.

OBJECTIVE: To determine the effectiveness of the unsubtracted roadmap technique (USRT) for decreasing cement leakage (CL) during percutaneous vertebroplasty (PVP). METHODS: In this retrospective cohort study, patients who underwent PVP between January and November 2018 were included. Patients were divided into the unsubtracted roadmap (UR) (n = 20) and conventional venography (CV) (n = 22) groups. USRT was performed before cement injection in the UR group. Computed tomography was performed on the first day after PVP in both groups to assess the CL. The vertebral height restoration rate was confirmed by radiography before and after the procedure. The CL location was classified as basivertebral vein (type B), segmental vein (type S), or cortical defect (type C). Visual analog scale and Oswestry Disability Index scores before and after surgery were calculated to confirm clinical outcomes. RESULTS: Based on the computed tomography images, CL occurred in 7 (34.78%) and 15 (63.64%) cases in the UR and CV groups, respectively. Number of CL type B, S, and C in the UR and CV groups were 2 and 6, 3 and 6, and 2 and 3, respectively. The injected cement volume was not significantly different between the UR (4.86 mL) and the CV (4.75 mL) groups (P = 0.450). Regarding vertebral body height restoration (P = 0.856) and clinical outcomes, there were no significant differences between both groups. Radiation exposure was significantly less in the UR group (P = 0.019). CONCLUSIONS: USRT is an effective method for reducing CL and radiation exposure during PVP.

Does Diabetes Affect the Surgical Outcomes in Cases With Cervical Ossification of the Posterior Longitudinal Ligament? A Multicenter Study From Asia Pacific Spine Study Group.

STUDY DESIGN: Retrospective multicenter study. OBJECTIVES: To evaluate the surgical outcomes of cervical ossification of the posterior longitudinal ligament (OPLL) in diabetes mellitus (DM) patients. METHODS: Approximately 253 cervical OPLL patients who underwent surgical decompression with or without fixation were registered at 4 institutions in 3 Asian countries. They were followed up for at least 2 years. Demographics, imaging, and surgical information were collected, and cervical Japanese Orthopaedic Association (JOA) scores and the visual analog scale (VAS) for the neck were used for evaluation. RESULTS: Forty-seven patients had DM, showing higher hypertension and cardiovascular disease prevalence. Although they presented worse preoperative JOA scores than non-DM patients (10.5 ± 3.1 vs. 11.8 ± 3.2; P = 0.01), the former showed comparable neurologic recovery at the final follow-up (13.9 ± 2.9 vs. 14.2 ± 2.6; P = 0.41). No correlation was noted between the hemoglobin A1c level in the DM group and the pre- and postoperative JOA scores. No significant difference was noted in VAS scores between the groups at pre- and postsurgery. Regarding perioperative complications, DM patients presented a higher C5 palsy frequency (14.9% vs. 5.8%; P = 0.04). A similar trend was observed when surgical procedure was limited to laminoplasty. CONCLUSIONS: This is the first multicenter Asian study to evaluate the impact of DM on cervical OPLL patients. Surgical results were favorable even in DM cases, regardless of preoperative hemoglobin A1c levels or operative procedures. However, caution is warranted for the occurrence of C5 palsy after surgery.

Association of frailty with regional sagittal spinal alignment in the elderly.

The degenerative changes in the spine of the frail elderly gradually exacerbate the alignment of the spine as the degeneration progresses. This study was conducted to assess the relationship between frailty and spine sagittal alignment measured in terms of global, cervical, thoracic, and lumbo-pelvic parameters. In total, 101 patients aged 75 years and older hospitalized for spine surgery were prospectively enrolled. We evaluated spinal sagittal parameters by dividing them into global (C7 sagittal vertical axis [SVA] and T1 pelvic angle [T1PA]), cervical (the C2-7 Cobb angle, Jackson line, and C2-7 plumb line), thoracic (thoracic kyphosis [TK]), and lumbo-pelvic (pelvic tilt [PT] and pelvic incidence minus lumbar lordosis value [PI-LL]). Patient characteristics; the Fatigue, Resistance, Ambulation, Illness, Loss of Weight (FRAIL) scale; and sagittal spinal parameters were included in the analysis. Multiple regression analysis was performed to identify associations between the FRAIL scale and sagittal spinal parameters. The FRAIL scale showed correlations with global sagittal parameters (C7 SVA [ß = 0.225, p = 0.029] and T1PA [ß = 0.273, p = 0.008]) and lumbo-pelvic parameters (PT [ß = 0.294, p = 0.004] and PI-LL [ß = 0.323, p = 0.001). Cervical and thoracic parameters were not directly associated with the FRAIL scale. LL and PI-LL were associated with TK, and TK was associated with cervical parameters (the C2-7 Cobb angle, Jackson line and C2-7 plumb line). In conclusion, frailty status could be an important factor that influences sagittal spinal alignment in the elderly. In this study, it was found that frailty mainly affected the balance of lumbo-pelvic alignment, and consequently affected the balance of the whole spine.

DNA methylation-dependent regulation of TrkA, TrkB, and TrkC genes in human hepatocellular carcinoma.

The tropomyosin-related kinase (Trk) family of neurotrophin receptors, TrkA, TrkB and TrkC, has been implicated in the growth and survival of human cancers. Here we report that Trks are frequently overexpressed in hepatocellular carcinoma (HCC) from patients and human liver cancer cell lines. To unravel the underlying molecular mechanism(s) for this phenomenon, DNA methylation patterns of CpG islands in TrkA, TrkB, and TrkC genes were examined in normal and cancer cell lines derived from liver. A good correlation was observed between promoter hypermethylation and lower expression of TrkA, TrkB, and TrkC genes, which was supported by the data that inhibiting DNA methylation with 5-azacytidine restored expression of those genes in normal liver cell lines. Furthermore, Trks promoted the proliferation of HepG2 and induced expression of the metastatic regulator, Twist. These results suggest that Trks may contribute to growth and metastasis of liver cancer.

Accessible chromatin structure permits factors Sp1 and Sp3 to regulate human TGFBI gene expression.

Transforming growth factor beta 1-induced (TGFBI) protein is an extracellular matrix (ECM) protein that is associated with other ECM proteins and functions as a ligand for various types of integrins. In this study, we investigated how human TGFBI expression is regulated in lung and breast cancer cells. We observed that the TGFBI promoter in A549 and MBA-MD-231 cells, which constitutively express TGFBI, existed in an open chromatin conformation associated with transcriptionally permissive histone modifications. Moreover, we found that TGFBI expression required Sp1 transcription elements that can bind transcription factors Sp1 and Sp3 in vitro. Occupancy of the TGFBI promoter by Sp1 and Sp3 in vivo was only observed in TGFBI-expressing cells, indicating that open chromatin conformation might facilitate the binding of Sp1 and Sp3 to the TGFBI promoter region. TGFBI promoter activity was impaired when Sp1 elements were mutated, but was increased when Sp1 or Sp3 factors was overexpressed. Furthermore, Sp1 inhibition in vivo by mithramycin A, as well as knockdown of Sp1 and/or Sp3 expression by short interfering RNA, significantly reduced TGFBI mRNA and protein levels. Thus, our data demonstrated that the expression of TGFBI is well correlated with chromatin conformation at the TGFBI promoter, and that factors Sp1 and Sp3 are the primary determinants for the control of constitutive expression of TGFBI gene.

A study using a Monte Carlo method of the optimal configuration of a distribution network in terms of power loss sensing.

Recently there have been many studies of power systems with a focus on "New and Renewable Energy" as part of "New Growth Engine Industry" promoted by the Korean government. "New And Renewable Energy"-especially focused on wind energy, solar energy and fuel cells that will replace conventional fossil fuels-is a part of the Power-IT Sector which is the basis of the SmartGrid. A SmartGrid is a form of highly-efficient intelligent electricity network that allows interactivity (two-way communications) between suppliers and consumers by utilizing information technology in electricity production, transmission, distribution and consumption. The New and Renewable Energy Program has been driven with a goal to develop and spread through intensive studies, by public or private institutions, new and renewable energy which, unlike conventional systems, have been operated through connections with various kinds of distributed power generation systems. Considerable research on smart grids has been pursued in the United States and Europe. In the United States, a variety of research activities on the smart power grid have been conducted within EPRI's IntelliGrid research program. The European Union (EU), which represents Europe's Smart Grid policy, has focused on an expansion of distributed generation (decentralized generation) and power trade between countries with improved environmental protection. Thus, there is current emphasis on a need for studies that assesses the economic efficiency of such distributed generation systems. In this paper, based on the cost of distributed power generation capacity, calculations of the best profits obtainable were made by a Monte Carlo simulation. Monte Carlo simulations that rely on repeated random sampling to compute their results take into account the cost of electricity production, daily loads and the cost of sales and generate a result faster than mathematical computations. In addition, we have suggested the optimal design, which considers the distribution loss associated with power distribution systems focus on sensing aspect and distributed power generation.