Peripheral CCL2-CCR2 signalling contributes to chronic headache-related sensitization.

Migraine, especially chronic migraine, is highly debilitating and still lacks effective treatment. The persistent headache arises from activation and sensitization of primary afferent neurons in the trigeminovascular pathway, but the underlying mechanisms remain incompletely understood. Animal studies indicate that signalling through chemokine C-C motif ligand 2 (CCL2) and C-C motif chemokine receptor 2 (CCR2) mediates the development of chronic pain after tissue or nerve injury. Some migraine patients had elevated CCL2 levels in CSF or cranial periosteum. However, whether the CCL2-CCR2 signalling pathway contributes to chronic migraine is not clear. Here, we modelled chronic headache with repeated administration of nitroglycerin (NTG, a reliable migraine trigger in migraineurs) and found that both Ccl2 and Ccr2 mRNA were upregulated in dura and trigeminal ganglion (TG) tissues that are implicated in migraine pathophysiology. In Ccl2 and Ccr2 global knockout mice, repeated NTG administration did not evoke acute or persistent facial skin hypersensitivity as in wild-type mice. Intraperitoneal injection of CCL2 neutralizing antibodies inhibited chronic headache-related behaviours induced by repeated NTG administration and repetitive restraint stress, suggesting that the peripheral CCL2-CCR2 signalling mediates headache chronification. We found that CCL2 was mainly expressed in TG neurons and cells associated with dura blood vessels, whereas CCR2 was expressed in subsets of macrophages and T cells in TG and dura but not in TG neurons under both control and disease states. Deletion of Ccr2 gene in primary afferent neurons did not alter NTG-induced sensitization, but eliminating CCR2 expression in either T cells or myeloid cells abolished NTG-induced behaviours, indicating that both CCL2-CCR2 signalling in T cells and macrophages are required to establish chronic headache-related sensitization. At cellular level, repeated NTG administration increased the number of TG neurons that responded to calcitonin-gene-related peptide (CGRP) and pituitary adenylate cyclase activating polypeptide (PACAP) as well as the production of CGRP in wild-type but not Ccr2 global knockout mice. Lastly, co-administration of CCL2 and CGRP neutralizing antibodies was more effective in reversing NTG-induced behaviours than individual antibodies. Taken together, these results suggest that migraine triggers activate CCL2-CCR2 signalling in macrophages and T cells. This consequently enhances both CGRP and PACAP signalling in TG neurons, ultimately leading to persistent neuronal sensitization underlying chronic headache. Our work not only identifies the peripheral CCL2 and CCR2 as potential targets for chronic migraine therapy, but also provides proof-of-concept that inhibition of both peripheral CGRP and CCL2-CCR2 signalling is more effective than targeting either pathway alone.

Real-Time Moving Object Tracking on Smartphone Using Cradle Head Servo Motor.

The increasing demand for artificially intelligent smartphone cradles has prompted the need for real-time moving object detection. Real-time moving object tracking requires the development of algorithms for instant tracking analysis without delays. In particular, developing a system for smartphones should consider different operating systems and software development environments. Issues in current real-time moving object tracking systems arise when small and large objects coexist, causing the algorithm to prioritize larger objects or struggle with consistent tracking across varying scales. Fast object motion further complicates accurate tracking and leads to potential errors and misidentification. To address these issues, we propose a deep learning-based real-time moving object tracking system which provides an accuracy priority mode and a speed priority mode. The accuracy priority mode achieves a balance between the high accuracy and speed required in the smartphone environment. The speed priority mode optimizes the speed of inference to track fast-moving objects. The accuracy priority mode incorporates CSPNet with ResNet to maintain high accuracy, whereas the speed priority mode simplifies the complexity of the convolutional layer while maintaining accuracy. In our experiments, we evaluated both modes in terms of accuracy and speed.

Incidence and characteristics of cataract surgery in South Korea from 2011 to 2015: A nationwide population-based study.

IMPORTANCE: Detailed incidence data for cataract surgery in the general population are limited, yet important for determining the surgical needs of the community and formulation of healthcare policies. BACKGROUND: To report incidence rates of cataract surgery in South Korea. DESIGN: Nationwide, retrospective population-based study. PARTICIPANTS: This study involved the entire population of South Korea (n = 47 990 761); 2 236 107 eyes of 1 591 176 patients confirmed as having cataract surgery from 1 January 2011 to 31 December 2015 were included. METHODS: Data for all patients who underwent primary cataract surgery in South Korea were retrieved using Korean Electronic Data Interchange and Korean Standard Classification of Diseases-7 codes. Annual incidence rates were calculated and adjusted to the national population data for the corresponding year. MAIN OUTCOME MEASURES: The average incidence of cataract surgery during the 5-year study period was estimated using population data from the 2010 Korean census. RESULTS: The incidence of cataract surgery increased from 8.54/1000 person-years in 2011 to 9.67/1000 person-years in 2015. The probability of second-eye surgery within 12 months after the first-eye surgery increased from 42.98% in 2011 to 48.01% in 2015. In total, 85.72% of surgeries were performed in non-rural areas: 43.18% in individuals with a higher household income and 76.65% in primary healthcare centres. The rate of vitrectomy for posterior capsular rupture was 0.72%. CONCLUSIONS AND RELEVANCE: The incidence of cataract surgery in South Korea is increasing over time. Our findings are expected to aid in the formulation of future healthcare policies concerning cataract surgery in South Korea.

Incidence and risk factors for retinal detachment after cataract surgery in Korea: a nationwide population-based study from 2011 to 2015.

PURPOSE: To evaluate the incidence and risk factors for retinal detachment (RD) after cataract surgery in the years 2011 to 2015 in Korea. METHODS: A nationwide retrospective cohort study was performed using health claim data from the Korean National Health Insurance Service (KNHIS) database. Patients over 40 years of age who underwent cataract surgery from 2011 and 2015 in Korea were retrospectively identified using Korean Electronic Data Interchange (KEDI) code and Korean Classification of Diseases (KCD)-7 code. RESULTS: A total of 2,191,510 eyes in 1,455,968 patients (58.63% female; mean age, 69.19 ± 9.82 years) underwent cataract surgery from 2011 to 2015 in Korea and 17,351 patients experienced RD (45.4% female; mean age, 60.89 ± 10.21 years). The 5-year cumulative risk of RD after cataract surgery was 1.19%, and 80.9% of RD occurred within 1 year after cataract surgery. In multivariate analysis, adjusted hazard ratio (HR) of RD was 1.335 [95% confidence interval (CI), 1.293-1.378] for male gender, 1.422 [95% CI, 1.371-1.475] for preoperative myopia, and 2.596 [95% CI, 2.367-2.849] for anterior vitrectomy during cataract surgery. Younger age was one of the factors highly associated with RD after cataract surgery, with HR [95% CI], 5.873 [5.527-6.240] in 40 to 54 years of age, 4.037 [3.811-4.277] in 55 to 64 years, and 2.026 [1.911-2.147] in 65 to 74 years. Adjusted HR of RD for surgery in secondary and primary healthcare centers were 0.495 [95% CI, 0.477-0.513] and 0.108 [95% CI, 0.104-0.113], respectively. Residence in non-metropolitan area and lower household income was associated with higher risk of RD. CONCLUSIONS: Younger age, anterior vitrectomy for posterior capsule rupture, preoperative myopia, male gender, surgery in tertiary referral centers, residence in non-metropolitan area, and lower household income were associated with an increased risk of RD after cataract surgery. The optimal timing of cataract surgery should be determined considering patient's risk factors, and appropriate pre- and postoperative evaluation is needed to prevent RD in patients with higher risks.

Clinical and Cephalometric Analysis of Facial Soft Tissue.

OBJECTIVE: This study aimed to provide average of standard values in planning orthognathic surgery in Asians. MATERIALS AND METHODS: Thirty-three Asians with well-balanced facial profile, combined with class I occlusion and stabilized condylar head were evaluated using lateral cephalograms. RESULTS: Facial length (Nasion'-Menton') was 138.8 and 127.0 mm in male and female, respectively. Upper and lower lip length were 24.5 and 49.8 mm for male, and 22.2 and 45.1 mm for female, and maxillary incisor exposure was 2.0 and 4.0 mm in male and female, respectively. Nasolabial angle was 77.7° and 84.1° in male and female, respectively. Alar base, A point', and maxillary incisor were placed posteriorly to true vertical line by 10.6, 1.0, and 8.0 mm for male and 9.0, 0.8, and 6.9 mm for female. The horizontal distance between upper lip anterior and lower lip anterior was 2.1 mm for male and 2.6 mm for female, and the horizontal distance between A point' and B point' was 5.3 mm for male and 3.9 mm for female. Orbital rim' to A-point' was 12.4 and 11.3 mm in male and female, respectively. Pogonion' located posteriorly to glabella' by 2.7 mm for male and anteriorly to glabella' by 3.2 mm for female, and facial angle was 156.7° and 147.0° in male and female, respectively. CONCLUSIONS: This quantitative analysis of facial profile in Asian will be helpful in evaluation of facial soft tissue and establishment of treatment plans for orthognathic surgery.

The family-wide structure and function of human dual-specificity protein phosphatases.

Dual-specificity protein phosphatases (DUSPs), which dephosphorylate both phosphoserine/threonine and phosphotyrosine, play vital roles in immune activation, brain function and cell-growth signalling. A family-wide structural library of human DUSPs was constructed based on experimental structure determination supplemented with homology modelling. The catalytic domain of each individual DUSP has characteristic features in the active site and in surface-charge distribution, indicating substrate-interaction specificity. The active-site loop-to-strand switch occurs in a subtype-specific manner, indicating that the switch process is necessary for characteristic substrate interactions in the corresponding DUSPs. A comprehensive analysis of the activity-inhibition profile and active-site geometry of DUSPs revealed a novel role of the active-pocket structure in the substrate specificity of DUSPs. A structure-based analysis of redox responses indicated that the additional cysteine residues are important for the protection of enzyme activity. The family-wide structures of DUSPs form a basis for the understanding of phosphorylation-mediated signal transduction and the development of therapeutics.

Molecular characterization of manganese peroxidases from white-rot fungus Polyporus brumalis.

The cDNAs of six manganese-dependent peroxidases (MnPs) were isolated from white-rot fungus Polyporus brumalis. The MnP proteins shared similar properties with each other in terms of size (approximately 360-365 amino acids) and primary structure, showing 62-96 % amino acid sequence identity. RT-PCR analysis indicated that these six genes were predominantly expressed in shallow stationary culture (SSC) in a liquid medium. Gene expression was induced by treatment with dibutyl phthalate (DBP) and wood chips. Expression of pbmnp4 was strongly induced by both treatments, whereas that of pbmnp5 was induced only by DBP, while pbmnp6 was induced by wood chips only. Then, we overexpressed pbmnp4 in P. brumalis under the control of the GPD promoter. Overexpression of pbmnp4 effectively increased MnP activity; the transformant that had the highest MnP activity also demonstrated the most effective decolorization of Remazol Brilliant Blue R dye. Identification of MnP cDNAs can contribute to the efficient production of lignin-degradation enzymes and may lead to utilization of basidiomycetous fungi for degradation of lignin and numerous recalcitrant xenobiotics.

Risk of stroke associated with intravitreal ranibizumab injections in age-related macular degeneration: a nationwide case-crossover study.

OBJECTIVE: To evaluate the risk of stroke associated with intravitreal ranibizumab in age-related macular degeneration (AMD). METHODS: A nationwide retrospective case-crossover study was performed using data from the Korean National Health Insurance Service (KNHIS) database, which included patients with exudative AMD in South Korea (n = 41,860). The index date was the date of hospitalization for stroke. We defined the case period as 60 days and four control periods before the index date. A pharmacy prescription database was searched for ranibizumab use during the case and control periods. We calculated adjusted odds ratios (ORs) and 95% confidence intervals (CIs) with a conditional logistic regression model. RESULTS: A total of 865 patients with AMD and incident stroke were included. Of all the patients, 12.02% had been treated during the preceding 60-day case period, compared with 9.25-10.29% during control periods. The adjusted OR of stroke associated with intravitreal ranibizumab during the case period was 1.285 (95% CI 0.979-1.686) (p = 0.07). In the subgroup analysis, the risk of hemorrhagic stroke had an OR of 2.252 (95% CI 1.068-4.749, p = 0.033). Further analyses based on patient gender, age, and different risk periods of 15 and 30 days yielded no increase in the risk of stroke associated with intravitreal ranibizumab. CONCLUSIONS: This case-crossover analysis revealed no evidence of increased risk of hospitalization for stroke within 60 days of intravitreal ranibizumab injection in AMD patients. A secondary analysis indicated the possibility of an increased risk of hemorrhagic stroke, with borderline significance. Further research is needed regarding the underlying biological mechanisms and drug safety.

The Protective Effects of Dexmedetomidine Preconditioning on Hepatic Ischemia/Reperfusion Injury in Rats.

BACKGROUND: Ischemia/reperfusion (IR) injury is 1 of the major problems in liver surgery. This study aims to evaluate the histologic and biochemical effects of dexmedetomidine on ischemia/reperfusion injury in the liver of rats. METHODS: Twenty-two Sprague-Dawley male rats were separated into 3 groups: group sham, IR (IR injury), and IR-D (IR with dexmedetomidine). Ischemia was induced for 45 minutes with portal clampage and the reperfusion period was 120 minutes. Group IR-D received 3 µg/kg of dexmedetomidine with loading for 10 minutes and then 3 µg/kg/h of dexmedetomidine was continuously injected intravenously 30 minutes before portal clampage. Biochemical factors (alanine aminotransferase and aspartate aminotransferase), variable cytokines (B cell lymphoma-2 (Bcl-2), Bax, caspase 3, caspase 8, nuclear factor-kappa B, interleukin (IL)-1ß, IL-6, IL-10, mixed lineage kinase domain-like protein, and receptor-interacting protein kinase-3), and histologic findings were investigated. RESULTS: Dexmedetomidine preconditioning significantly suppressed the histologic damage. In the IR-D group, the expression of IL-6 was decreased and the Bcl-2 was increased when compared with the IR group. CONCLUSION: Dexmedetomidine suppresses hepatic IR injury and the protective mechanism appears to involve the decrease of IL-6 and upregulation of Bcl-2 expression, which result in the attenuation of inflammatory response and the inhibition of apoptosis.

Oncogenic Ras-mediated downregulation of Clast1/LR8 is involved in Ras-mediated neoplastic transformation and tumorigenesis in NIH3T3 cells.

Oncogenic Ras proteins transform cells by way of multiple downstream signaling pathways that promote the genesis of human cancers. However, the exact cellular mechanisms by which downstream targets are regulated are not fully understood. Here, we show that oncogenic Ras reduced Clast1/LR8 transcript levels in mouse NIH3T3 fibroblasts and human WI38 fibroblasts. Clast1/LR8 transcript was undetectable in H460, A549, and H1299 cells showing high Ras activity, but was relatively abundant in DMS53 cells displaying low Ras activity. We also showed that K-Ras siRNA restored Clast1/LR8 expression in H460 and A549 cells, and that inhibitors of DNA methylation and histone deacetylation reversed oncogenic H-Ras-mediated suppression of Clast1/LR8 transcription. Additionally, ectopic expression of Clast1/LR8 inhibited serum-stimulated phosphorylation of ERK1/2 and Akt in H-RasV12-transformed NIH3T3 cells. We further showed that the expression of Clast1/LR8 interfered with oncogenic Ras-induced NIH3T3 cell transformation and invasion. Finally, our results showed that Clast1/LR8 inhibited Ras-induced proliferation of, and tumor formation by, oncogenic H-RasV12-transformed NIH3T3 cells in vivo. This study identifies the downregulation of Clast1/LR8 as a potentially important mechanism by which oncogenic Ras-mediated neoplastic transformation occurs.

Implantation of polymer scaffolds seeded with neural stem cells in a canine spinal cord injury model.

BACKGROUND AIMS: Combinatorial approaches employing diverse therapeutic modalities are required for clinically relevant repair of injured spinal cord in human patients. Before translation into the clinic, the feasibility and therapeutic potential of such combinatorial strategies in larger animal species need to be examined. METHODS: The present study tested the feasibility of implanting polymer scaffolds via neural stem cell (NSC) delivery in a canine spinal cord injury (SCI) model. The poly(lactic-co-glycolic acid) (PLGA) scaffolds seeded with human NSC were implanted into hemisected canine spinal cord. RESULTS: The PLGA scaffolds bridged tissue defects and were nicely integrated with residual canine spinal cord tissue. Grafted NSC survived the implantation procedure and showed migratory behavior to residual spinal cord tissue. Ectopic expression of a therapeutic neurotrophin-3 gene was also possible in NSC seeded within the PLGA scaffolds. CONCLUSIONS: Our description of a canine SCI model would be a valuable resource for pre-clinical trials of combinatorial strategies in larger animal models.

Effects of installation depth on survival of an hydroxyapatite-coated Bicon implant for single-tooth restoration.

PURPOSE: The present retrospective study evaluated various implant surface factors associated with Bicon implant survival for single-tooth restoration in the healthy individual. PATIENTS AND METHODS: A retrospective cohort study design was used. A total of 613 Bicon (Bicon System, Boston, MA) implants (272 patients) were included. Because the use of hydroxyapatite (HA) coating has been controversial, the surface type was chosen according to the patient's preference. A total of 308 HA-coated implants and 305 titanium plasma-sprayed (TPS) implants were used. The macroanatomy of both implant types was identical. Patients who had systemic disease, poor quality bone, or removable prosthetics were excluded from the present study. A chart review was conducted to record age, gender, implant diameter, implant length, installation depth, installation location, and the use of bone grafting. Implant failure was recorded, and the data were analyzed using the chi(2) test and logistic regression analysis. RESULTS: The installation depth was an important prognostic factor in the HA-coated implants. The failure rate for the HA-coated implants installed at margin level and 2 mm below level was 10.29% and 3.01%, respectively (chi(2) = 6.035, P = .014). The implant length was an important prognostic factor for the TPS-treated implants. The failure rate recorded for the TPS-treated implants installed with a length of less than 10 mm and 10 mm or longer was 15.46% and 2.40%, respectively (chi(2) = 18.414, P < .001). CONCLUSION: Many factors can influence the failure rate of TPS and HA-coated Bicon implants. Among these, installation depth played an especially significant role in the success of HA-coated Bicon implants in the present study.

Association between Ranibizumab Injections and Risk of Acute Myocardial Infarction in Age-related Macular Degeneration: A Case-crossover Study.

PURPOSE: This study aimed to evaluate the risk of acute myocardial infarction (AMI) associated with intravitreal ranibizumab in age-related macular degeneration (AMD). METHODS: This nationwide retrospective case-crossover study using data from the Korean National Health Insurance Service database included patients diagnosed with exudative AMD using the registration code for exudative AMD (V201) from 2009 to 2014. We identified all incident AMI cases among these exudative AMD cases from inpatient claims and defined the index date as the date of hospitalization. For each patient, we defined the case period as one to 60 days and four control periods as 121 to 180, 181 to 240, 241 to 300, and 301 to 361 days, respectively, before the index date. A prescription of ranibizumab was searched for during the case and control periods. We calculated the adjusted odds ratios and their 95% confidence intervals using a conditional logistic regression model. RESULTS: From a cohort of patients with exudative AMD (n = 41,860), a total of 181 AMI patients with exudative AMD were included. Among all the patients, 11.05% were treated during the 2 months preceding the index date as compared with 8.29% to 9.39% treated during control periods. The adjusted odds ratio of AMI associated with intravitreal ranibizumab during the preceding 2 months was 1.22 (95% confidence interval, 0.673-2.213; p = 0.5124). Analyses based on case periods of 15 days and 1 month yielded similar results. CONCLUSIONS: Intravitreal ranibizumab injection does not appear to increase the risk of hospitalization for AMI within 60 days in exudative AMD patients.

Korean red ginseng promotes hippocampal neurogenesis in mice.

Neurogenesis in the adult hippocampus plays a major role in cognitive ability of animals including learning and memory. Korean red ginseng (KRG) has long been known as a medicinal herb with the potential to improve learning and memory; however, the mechanisms are still elusive. Therefore, we evaluated whether KRG can promote cognitive function and enhance neurogenesis in the hippocampus. Eight-week-old male C57BL/6 mice received 50 mg/kg of 5-bromo-2'-deoxyuridine (BrdU) intraperitoneally and 100 mg/kg of KRG or vehicle orally once a day for 14 days. Pole, Rotarod and Morris water maze tests were performed and the brains were collected after the last behavioral test. Changes in the numbers of BrdU- and BrdU/doublecortin (DCX; a marker for neuronal precursor cells and immature neurons)-positive cells in the dentate gyrus and the gene expression of proliferating cell nuclear antigen (a marker for cell differentiation), cerebral dopamine neurotrophic factor and ciliary neurotrophic factor in the hippocampus were then investigated. KRG-treated mice came down the pole significantly faster and stood on the rotarod longer than vehicle-treated mice. The Morris water maze test showed that KRG administration enhanced the learning and memory abilities significantly. KRG also significantly increased BrdU- and BrdU/DCX-positive cells in the dentate gyrus as well as the proliferating cell nuclear antigen, cerebral dopamine neurotrophic factor and ciliary neurotrophic factor mRNA expression levels in the hippocampus compared to vehicle. Administration of KRG promotes learning and memory abilities, possibly by enhancing hippocampal neurogenesis. This study was approved by the Pusan National University Institutional Animal Care and Use Committee (approval No. PNU-2016-1071) on January 19, 2016.

Ischemic Preconditioning Protects Against Hepatic Ischemia-Reperfusion Injury Under Propofol Anesthesia in Rats.

BACKGROUND: Propofol is widely used in general anesthesia, and it has been reported to protect various organs against ischemia-reperfusion injury (IRI), including liver. To evaluate the hepatoprotective effects of ischemic preconditioning (IP) under propofol anesthesia, we investigated the possible underlying mechanisms in rats. METHODS: Male Sprague-Dawley rats were randomly assigned to 3 groups: sham group (n = 5), non-IP group (n = 9; 45 minutes of hepatic ischemia followed by 2 hours of reperfusion), and IP group (n = 9; IP applied as 10 minutes of hepatic ischemia followed by 15 minutes of reperfusion before 45 minutes of ischemia). Anesthesia was maintained with intravenous (IV) infusion of propofol (800 µg/kg/min). Liver enzymes, histopathological changes, and cytokine expression were examined. RESULTS: The IP group showed significantly lower liver enzyme levels (aspartate aminotransferase, P = .045; alanine aminotransferase, P = .006) and reduced the histologic grades of hepatic injury 2 hours after reperfusion (P = .004) compared to the non-IP group. Lactate dehydrogenase activity (P < .001) and interleukin-6 mRNA levels were significantly higher in the non-IP group than in the sham and IP groups (P = .002, both groups). CONCLUSIONS: Our results demonstrate that IP under propofol anesthesia significantly attenuated hepatic IRI. The principal mechanism of the protective effects appeared to involve reduced expression of the IL-6 pro-inflammatory cytokine and subsequent reduction of the degree of necrosis.

Molecular characterization of the sweet potato peroxidase SWPA4 promoter which responds to abiotic stresses and pathogen infection.

Previously, the swpa4 peroxidase gene has been shown to be inducible by a variety of abiotic stresses and pathogenic infections in sweet potato (Ipomoea batatas). To elucidate its regulatory mechanism at the transcriptional level under various stress conditions, we isolated and characterized the promoter region (2374 bp) of swpa4 (referred to as SWPA4). We performed a transient expression assay in tobacco protoplasts with deletions from the 5'-end of SWPA4 promoter fused to the beta-glucuronidase (GUS) reporter gene. The -1408 and -374 bp deletions relative to the transcription start site (+1) showed 8 and 4.5 times higher GUS expression than the cauliflower mosaic virus 35S promoter, respectively. In addition, transgenic tobacco plants expressing GUS under the control of -2374, -1408 or -374 bp region of SWPA4 promoter were generated and studied in various tissues under abiotic stresses and pathogen infection. Gel mobility shift assays revealed that nuclear proteins from sweet potato cultured cells specifically interacted with 60-bp fragment (-178/-118) in -374 bp promoter region. In silico analysis indicated that four kinds of cis-acting regulatory sequences, reactive oxygen species-related element activator protein 1 (AP1), CCAAT/enhancer-binding protein alpha element, ethylene-responsive element (ERE) and heat-shock element, are present in the -60 bp region (-178/-118), suggesting that the -60 bp region might be associated with stress inducibility of the SWPA4 promoter.

Differential expression of HSP70 mRNA in the mouse brain after treatment with geldanamycin.

OBJECTIVES: The effect of geldanamycin (GA) on the brain is poorly understood. The induction of specific heat shock protein (HSP) isoforms by GA has not been studied in detail. The aim of this work was to identify the HSP70 genes that become up-regulated in the mouse brain after GA administration. METHODS: The brains of male hsp 70.1 knockout and littermate mice were obtained after GA injection into the lateral ventricle. The mice were killed 6 hours later to investigate the levels of hsp 70.1 and hsp 70.3 using semi-quantitative reverse transcriptase polymerase chain reactions. RESULTS: GA infusion into the mouse brain led to the up-regulation of hsp 70.1. DISCUSSION: Further work is needed to more clearly define the role of those genes in the clinical effects of GA.

Therapeutic efficacy of neuregulin 1-expressing human adipose-derived mesenchymal stem cells for ischemic stroke.

Adipose-derived mesenchymal stem cells (AdMSCs) have been reported to ameliorate neurological deficits after acute ischemic stroke. As neuregulin 1 (NRG1, or heregulin 1), a growth factor with versatile functions in the central nervous system, has demonstrated protective effects against ischemic brain injuries, we have generated NRG1-overexpressing AdMSCs in order to investigate whether NRG1-AdMSCs could enhance therapeutic benefits of AdMSCs in ischemic stroke. After AdMSCs were infected with adenoviral NRG1, increased NRG1 secretion in NRG1-AdMSCs was confirmed with ELISA. At 1 d after ischemic stroke that was induced by the occlusion of middle cerebral artery (MCAo) for 60 min in Sprague Dawley (SD) rats, adenoviral NRG1, AdMSCs, NRG1-AdMSCs, or PBS were injected into the striatum and serial neurologic examinations were performed. Administration of NRG1-AdMSCs resulted in significant improvement of functional outcome following stroke compared to AdMSCs- or adenoviral NRG1-treated group, in addition to the reduction in the infarct size evaluated by hematoxylin and eosin staining. When NRG1 expression in the brain was examined by double immunofluorescence to human nuclei (HuNu)/NRG1 and ELISA, NRG1-AdMSCs demonstrated marked increase in NRG1 expression. Moreover, western blot analysis further showed that transplantation of NRG1-AdMSCs significantly increased both endogenous and adenoviral NRG1 expression compared to AdMSCs-treated group. To elucidate molecular mechanisms, NRG1-associated downstream molecules were evaluated by western blot analysis. Expression of ErbB4, a receptor for NRG1, was markedly increased by NRG1-AdMSCs administration, in addition to pMAPK and pAkt, crucial molecules of NRG1-ErbB4 signaling. Taken together, our data suggest that NRG1-AdMSCs can provide excellent therapeutic potential in ischemic stroke by activating NRG1-ErbB4 signaling network.

A principled strategy for mapping enhancers to genes.

Mapping enhancers to genes is a fundamental goal of modern biology. We have developed an innovative strategy that maps enhancers to genes in a principled manner. We illustrate its power by applying it to Myrf. Despite being a master regulator of oligodendrocytes, oligodendrocyte enhancers governing Myrf expression remain elusive. Since chromatin conformation capture studies have shown that a gene and its enhancer tend to be found in the same topologically associating domain (TAD), we started with the delineation of the Myrf TAD. A genome-wide map of putative oligodendrocyte enhancers uncovered 6 putative oligodendrocyte enhancers in the Myrf TAD, narrowing down the search space for Myrf enhancers from the entire genome to 6 loci in a principled manner. Epigenome editing experiments revealed that two of them govern Myrf expression for oligodendrocyte development. Our new method is simple, principled, and powerful, providing a systematic way to find enhancers that regulate the expression of a gene of interest. Since it can be applied to most cell types, it would greatly facilitate our effort to unravel transcriptional regulatory networks of diverse cell types.

Changes in Human Tear Proteome Following Topical Treatment of Dry Eye Disease: Cyclosporine A Versus Diquafosol Tetrasodium.

Purpose: To compare the changes in human tear proteome and clinical effects following topical cyclosporine A (CsA) 0.05% or diquafosol tetrasodium (DQS) 3% treatment of dry eye disease (DED), and to identify biomarkers for determining disease severity and treatment effectiveness in DED. Methods: A total of 18 patients were diagnosed with non-Sjögren DED. Nine patients in each group were treated with topical CsA 0.05% or DQS 3% for 4 weeks. Tear samples were collected after evaluation of tear breakup time, corneal and conjunctival erosion staining, and results of Schirmer's test 1 before and after treatment. Proteomes were characterized using liquid chromatography mass spectrometry, and proteins exhibiting a fold change >1.5 or <0.67 (P < 0.05) were considered differentially expressed (DEP). Results: A total of 794 proteins were identified, with no significant difference observed between pretreatment and posttreatment conditions. Proteomic analysis identified 54 and 106 DEPs between treatment groups (CsA and DQS, respectively), with gene ontology analysis indicating that both treatments enhanced innate and adaptive immune responses and cellular detoxification. Protein-network analysis showed that inflammation associated with the immune response was primarily responsible for the therapeutic process in both groups. Conclusions: These results provide insight into the broad scope of changes at the ocular surface in DED and indicated that although both drugs improved the clinical parameters, the activated tear-specific biomarkers differed significantly between treatments. Our findings suggest that the DEPs identified here and those correlated with the clinical parameters might represent candidate biomarkers for DED.