Cancer-associated fibroblasts induce an aggressive phenotypic shift in non-malignant breast epithelial cells via interleukin-8 and S100A8.

Cancer-associated fibroblasts (CAFs) in the tumor microenvironment have been associated with tumor progression in breast cancer. Although crosstalk between breast cancer cells and CAFs has been studied, the effect of CAFs on non-neoplastic breast epithelial cells is not fully understood to date. Here, we investigated the effect of CAFs on aggressive phenotypes in non-neoplastic MCF10A breast epithelial cells. CAFs induced epithelial-to-mesenchymal transition (EMT) and invasive phenotype in MCF10A cells. S100A8, a potential prognostic marker in several cancers, was markedly increased in MCF10A cells by CAFs. S100A8 was crucial for CAFs-induced invasive phenotype of MCF10A cells. Among cytokines increased by CAFs, interleukin (IL)-8 induced S100A8 through transcription factors p65 NF-κB and C/EBPß. In a xenograft mouse model with MCF10A cells and CAFs, tumor was not developed, suggesting that coinjection with CAFs may not be sufficient for in vivo tumorigenicity of MCF10A cells. Xenograft mouse tumor models with MDA-MB-231 breast carcinoma cells provided an in vivo evidence for the effect of CAFs on breast cancer progression as well as a crucial role of IL-8 in tumor growth and S100A8 expression in vivo. Using a tissue microarray of human breast cancer, we showed that S100A8 expression was correlated with poor outcomes. S100A8 expression was more frequently detected in cancer-adjacent normal human breast tissues than in normal breast tissues. Together, this study elucidated a novel mechanism for the acquisition of invasive phenotype of non-neoplastic breast cells induced by CAFs, suggesting that targeting IL-8 and S100A8 may be an effective strategy against breast cancer.

Optical frequency switching scheme for a high-speed broadband THz measurement system based on the photomixing technique.

This study presents an optical frequency switching scheme for a high-speed broadband terahertz (THz) measurement system based on the photomixing technique. The proposed system can achieve high-speed broadband THz measurements using narrow optical frequency scanning of a tunable laser source combined with a wavelength-switchable laser source. In addition, this scheme can provide a larger output power of an individual THz signal compared with that of a multi-mode THz signal generated by multiple CW laser sources. A swept-source THz tomography system implemented with a two-channel wavelength-switchable laser source achieves a reduced time for acquisition of a point spread function and a higher depth resolution in the same amount of measurement time compared with a system with a single optical source.

Continuous-wave THz vector imaging system utilizing two-tone signal generation and self-mixing detection.

We propose and demonstrate a continuous-wave vector THz imaging system utilizing a photonic generation of two-tone THz signals and self-mixing detection. The proposed system measures amplitude and phase information simultaneously without the local oscillator reference or phase rotation scheme that is required for heterodyne or homodyne detection. In addition, 2π phase ambiguity that occurs when the sample is thicker than the wavelength of THz radiation can be avoided. In this work, THz signal having two frequency components was generated with a uni-traveling-carrier photodiode and electro-optic modulator on the emitter side and detected with a Schottky barrier diode detector used as a self-mixer on the receiver side. The proposed THz vector imaging system exhibited a 50-dB signal to noise ratio and 0.012-rad phase fluctuation with 100-µs integration time at 325-GHz. With the system, we demonstrate two-dimensional THz phase contrast imaging. Considering the recent use of two-dimensional arrays of Schottky barrier diodes as a THz image sensor, the proposed system is greatly advantageous for realizing a real-time THz vector imaging system due to its simple receiver configuration.

The Novel Scoring System for 30-Day Mortality in Patients with Non-variceal Upper Gastrointestinal Bleeding.

BACKGROUND: Although the mortality rates for non-variceal upper gastrointestinal bleeding (NVUGIB) have recently decreased, it remains a significant medical problem. AIM: The main aim of this prospective multicenter database study was to construct a clinically useful predictive scoring system by using our predictors and compare its prognostic accuracy with that of the Rockall scoring system. METHODS: Data were collected from consecutive patients with NVUGIB. Logistic regression analysis was performed to identify the independent predictors of 30-day mortality. Each independent predictor was assigned an integral point proportional to the odds ratio (OR) and we used the area under the curve to compare the discrimination ability between the new predictive model and the Rockall score. RESULTS: The independent predictors of mortality included age >65 years [OR 2.627; 95 % confidence interval (CI) 1.298-5.318], hemodynamic instability (OR 2.217; 95 % CI 1.069-4.597), serum blood urea nitrogen level >40 mg/dL (OR 1.895; 95 % CI 1.029-3.490), active bleeding at endoscopy (OR 2.434; 95 % CI 1.283-4.616), transfusions (OR 3.811; 95 % CI 1.640-8.857), comorbidities (OR 3.481; 95 % CI 1.405-8.624), and rebleeding (OR 10.581; 95 % CI 5.590-20.030). The new predictive model showed a high discrimination capability and was significantly superior to the Rockall score in predicting the risk of death (OR 0.837;95 % CI 0.818-0.855 vs. 0.761; 0.739-0.782; P = 0.0123). CONCLUSIONS: The new predictive score was significantly more accurate than the Rockall score in predicting death in NVUGIB patients. We need to prospectively validate the accuracy of this score for predicting mortality in NVUGIB patients.

Sphingosine 1-phosphate regulates matrix metalloproteinase-9 expression and breast cell invasion through S1P3-Gαq coupling.

Recent evidence suggests that inflammation is involved in malignant progression of breast cancer. Sphingosine 1-phosphate (S1P), acting on the G-protein-coupled receptors, is known as a potent inflammatory mediator. In this study, the effect of the inflammatory lipid S1P on the regulation of invasive/migratory phenotypes of MCF10A human breast epithelial cells was investigated to elucidate a causal relationship between inflammation and the control of invasiveness of breast cells. We show that S1P causes induction of matrix metalloproteinase-9 (MMP-9) in vitro and in vivo, and thus enhances invasion and migration. We also show that fos plays a crucial role in the transcriptional activation of MMP-9 by S1P. In addition, activation of extracellular-signal-regulated kinases 1 and 2 (ERK1/2), p38 and alpha serine/threonine-protein kinase (Akt) are involved in the process of S1P-mediated induction of MMP-9 expression and invasion. Activation of the S1P receptor S1P3 and G(αq) are required for S1P-induced invasive/migratory responses, suggesting that the enhancement of S1P-mediated invasiveness is triggered by the specific coupling of S1P3 to the heterotrimeric G(αq) subunit. Activation of phospholipase C-ß4 and intracellular Ca²âº release are required for S1P-induced MMP-9 upregulation. Taken together, this study demonstrated that S1P regulates MMP-9 induction and invasiveness through coupling of S1P3 and G(αq) in MCF10A cells, thus providing a molecular basis for the crucial role of S1P in promoting breast cell invasion.

Attenuation of spinal cord injury-induced astroglial and microglial activation by repetitive transcranial magnetic stimulation in rats.

Spinal cord injury (SCI) causes not only loss of sensory and motor function below the level of injury but also chronic pain, which is difficult and challenging of the treatment. Repetitive transcranial magnetic stimulation (rTMS) to the motor cortex, of non-invasive therapeutic methods, has the motor and sensory consequences and modulates pain in SCI-patients. In the present study, we studied the effectiveness of rTMS and the relationship between the modulation of pain and the changes of neuroglial expression in the spinal cord using a rat SCI-induced pain model. Elevated expressions of Iba1 and GFAP, specific microglial and astrocyte markers, was respectively observed in dorsal and ventral horns at the L4 and L5 levels in SCI rats. But in SCI rats treated with 25 Hz rTMS for 8 weeks, these expressions were significantly reduced by about 30%. Our finding suggests that this attenuation of activation by rTMS is related to pain modulation after SCI. Therefore, rTMS might provide an alternative means of attenuating neuropathic pain below the level of SCI.

Pulsed radiofrequency application reduced mechanical hypersensitivity and microglial expression in neuropathic pain model.

OBJECTIVE: Pulsed radiofrequency (PRF) procedure has been used in clinical practice for the treatment of chronic neuropathic pain conditions without neuronal damage. The purpose of this study was to investigate the changes in pain response and glial expression after the application of PRF on a dorsal root ganglion (DRG) in a neuropathic pain model. DESIGN: A neuropathic pain model (14 female Sprague-Dawley [SD] rats; 200-250 g) was made by a unilateral L5 spinal nerve ligation (SNL) and transection on the distal side of the ligation. The development of mechanical and cold hypersensitivity on the hindpaw was established postoperative day 9 (POD 9). The rats were then randomly assigned to the PRF (+) and the PRF (-) groups. Furthermore, PRF (2 bursts/s, duration = 20 milliseconds, output voltage = 45 V) was applied on the ipsilateral DRG for 180 seconds, with a maximum temperature of 42°C, at POD 10. Pain behaviors were tested throughout the 12 days after PRF. We also examined the changes of the spinal glial expression by immunohistochemistry. RESULTS: Significant reduction of mechanical hypersensitivity in the PRF (+) group was observed from day 1 after a single PRF procedure and was maintained throughout the following 12 days. Immunoreactivity for OX42 in the ipsilateral dorsal horn also decreased compared with that of the PRF (-) group. However, cold hypersensitivity and glial fibrillary acidic protein (GFAP) immunoreactivity in the dorsal horn was not affected by a PRF procedure. CONCLUSIONS: Our result demonstrated that the mechanical hypersensitivity, induced by L5 SNL, was attenuated by a PRF procedure on the ipsilateral DRG. This analgesic effect may be associated with an attenuation of the microglial activation in the dorsal horn.

Mental Stress Assessment Using Ultra Short Term HRV Analysis Based on Non-Linear Method.

Mental stress is on the rise as one of the major health problems in modern society. It is important to detect and manage mental stress to prevent various diseases caused by stress and to maintain a healthy life. The purpose of this paper is to present new heart rate variability (HRV) features based on empirical mode decomposition and to detect acute mental stress through short-term HRV (5 min) and ultra-short-term HRV (under 5 min) analysis. HRV signals were acquired from 74 young police officers using acute stressors, including the Trier Social Stress Test and horror movie viewing, and a total of 26 features, including the proposed IMF energy features and general HRV features, were extracted. A support vector machine (SVM) classification model is used to classify the stress and non-stress states through leave-one-subject-out cross-validation. The classification accuracies of short-term HRV and ultra-short-term HRV analysis are 86.5% and 90.5%, respectively. In the results of ultra-short-term HRV analysis using various time lengths, we suggest the optimal duration to detect mental stress, which can be applied to wearable devices or healthcare systems.

A toenail flap based on the fibro-osseous hiatus branch for fingernail reconstruction.

This study included two parts: 1) cadaver dissection to elucidate the perfusion of toenail flaps by the fibro-osseous hiatus branch (FHB), and 2) clinical application of the toenail flap for reconstruction of a fingernail defect. Four second toes of two fresh Korean cadavers were dissected. The plantar digital artery (PDA) and terminal segment branch (TSB) were ligated, and red latex was injected distally into the ligated PDA. Perfusion of the dye into the toenail bed through the FHB was observed. From Oct 2004 to Sep 2009, eight toenail flaps based on the FHB pedicle with or without the distal phalanx and pulp were applied to seven patients for finger nail reconstruction. The toenail flap was marked at 5 mm distal to the nail fold and 5 mm lateral to the paronychium. The toenail complex based on the FHB was elevated and transferred to the finger. The nail and matrix were elevated with or without including the distal phalanx. The results of perfusion study showed that one side of the unilateral FHB was identified and traced proximal to the PDA, which was ligated. The distal toenail bed was perfused by the dye through the FHB. In clinical application, all the toenail flaps flourished and survived. We suggest that the toenail flap based on the FHB may be useful for fingernail reconstruction with minimal donor morbidity.

The expression of corticotropin-releasing factor in the central nucleus of the amygdala, induced by colorectal distension, is attenuated by general anesthesia.

Corticotrophin-releasing factor (CRF), a key regulator of the hypothalamic-pituitary axis, is expressed in the central nucleus of the amygdala (CeA) and its expression is upregulated in stress-related disorders. We investigated here the effect of noxious colorectal distension (CRD) on the expression of CRF in the CeA of conscious and unconscious rats. Adult male rats with or without general anesthesia were exposed to visceral pain induced by CRD for 5 min; this procedure was repeated 3 times with 1 min resting after each distension. The rats were sacrificed and sections of the CeA were immunostained for CRF as an indicator for anxiety response, and for phosphorylated extracellular signal-regulated kinase (p-ERK) as a marker for pain-specific activation of neurons; sections of lumbosacral spinal cord were immunostained for c-Fos as a marker for activation of spinal neurons. CRD elicited a significant increase in the expression of CRF and p-ERK in the CeA and of c-Fos in the spinal cord. General anesthesia attenuated the increase in CRF and p-ERK in the CeA, but did not affect the expression of spinal c-Fos. These results suggest that conscious recognition of pain at higher brain centers is an important determinant of CRF expression in the CeA.

Arterial anatomy of the second toe nail bed related to toenail transfer.

The aim of this study was to elucidate the exact course of the terminal branches of the plantar digital artery (PDA) to the nail bed of the second toe. Thirteen second toes from seven fresh Korean cadavers were dissected (age range 74-92 years, four men and three women). The terminal segmental branches (TSB) branched off from the PDA at 7.6 ± 0.7 mm proximal to the nail fold. The fibro-osseous hiatus branch (FHB) branched off from the PDA at 3.3 ± 0.7 mm from the nail fold. They were 3.8 ± 1.0 mm lateral to the paronychium. Diameters of TSB and FHB were 0.8 ± 0.2 mm and 0.7 ± 0.1 mm, respectively. Diameter of PDA was 1.4 ± 0.2 mm. Surgeons should stay at least 4 mm proximal to the nail fold to avoid injury to the terminal branch. We believe that second toenail with minimum amount of soft tissue may be transferred using FHB-based vascularized toenail flap. Perfusion study and clinical application should be followed.

The Effect of a Modified Constant Flow Insufflation of Oxygen during Cardiopulmonary Resuscitation in a Rat Model of Respiratory Cardiac Arrest on Arterial Oxygenation, Alveolar Barotrauma, and Brain Tissue Injury.

AIM: Intermittent positive pressure ventilation (IPPV) can adversely affect cardiopulmonary resuscitation outcomes by increasing the intrathoracic pressure. Continuous flow insufflation of oxygen (CFIO) has been investigated as a potential alternative, but evidence supporting its superiority over intermittent positive pressure ventilation in cases of cardiac arrest is scant. The aim of the current study was to compare the effects of continuous flow insufflation of oxygen using a one-way valve during cardiopulmonary-resuscitation with intermittent positive pressure ventilation in a rat model of respiratory arrest. METHODS: Male Sprague-Dawley rats weighing 400∼450 g (from minimum to maximum) were randomly assigned to either a sham, IPPV, or CFIO group (n = 10 per group). Respiratory arrest was induced by blocking the endotracheal tube. Arterial blood gas analysis was performed during cardiopulmonary resuscitation to compare the oxygenation levels. Tissues were then harvested to compare the degrees of pulmonary barotrauma and ischemic brain injury. RESULTS: Return of spontaneous circulation was observed in 6/10 rats in the IPPV group and 5/10 in the CFIO group. During cardiopulmonary resuscitation, the mean PaO2 was significantly higher in the CFIO group (83.10 mmHg) than in the IPPV group (56.10 mmHg). Lung biopsy revealed more inflammatory cells and marked thickening of the alveolar wall in the IPPV group; the group also exhibited a higher frequency of neuroglial cells and apoptotic bodies of pyramidal cells, resulting from ischemic injury. CONCLUSION: In a rat model of respiratory arrest, CFIO using a one-way valve resulted in a greater level of oxygenation and less lung and brain injuries than with IPPV.

Oral administration of 1,4-aryl-2-mercaptoimidazole inhibits T-cell proliferation and reduces clinical severity in the murine experimental autoimmune encephalomyelitis model.

T cells play a pivotal role in the initiation and progression of multiple sclerosis. We have found that 1,4-aryl-2-mercaptoimidazole (KRM-III) inhibited T-cell antigen receptor- and phorbol myristate acetate/ionomycin-induced activation of nuclear factor of activated T cells (NFAT) and T-cell proliferation with an IC(50) of 5 microM. The KRM-III-mediated inhibitory effect was specific for NFAT activation but not for nuclear factor kappaB. Oral administration of 90 mg/kg KRM-III resulted in complete abrogation of anti-CD3 antibody-induced T-cell activation and a 45.8% reduction in footpad swelling in bovine serum albumin-induced delayed-type hypersensitivity. In the murine experimental autoimmune encephalomyelitis (EAE) model, oral administration of KRM-III significantly attenuated the severity of disease when given before or after disease onset. Draining lymph node cells from KRM-III-treated mice showed markedly reduced proliferation in response to myelin oligodendrocyte glycoprotein peptide. Histological analysis indicated that KRM-III reduced the infiltration of inflammatory cells to the white matter of spinal lumbar cords. These results demonstrate that KRM-III efficiently inhibits T-cell activation and inflammatory responses and lessens EAE clinical signs, which suggest KRM-III as a potential lead compound for the treatment of T-cell-driven autoimmune diseases.

Proinflammatory cytokine and nitric oxide production by human macrophages stimulated with Trichomonas vaginalis.

Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.

Experimentally derived viscoelastic properties of human skin and muscle in vitro.

Measurement of the mechanical properties of human skin in vivo is challenging. Moreover, those with regard to excitation frequency have been rarely reported thus far. In this study, a vibration-based experimental method was employed to measure the viscoelastic properties with regard to the excitation frequency. Pieces of human skin and skeletal muscle excised from cadavers immediately post mortem were stored in a sealed container. As the experiment began, they were removed from the container and used to measure the viscoelastic properties as time elapsed. Young's moduli of the samples of human skin tissue that were immediately removed from the container were found to be similar to those obtained with in-vivo indentation methods. They were also found to be approximately one-third of those of human skeletal muscles. The viscoelastic properties of human skin were found to remain almost constant within the frequency range up to 120 Hz and are similar to those of porcine tissue. Young's moduli of the human skin and skeletal muscle were also found to reach the maximum values approximately five days post mortem. However, the loss factor of the human skin did not vary significantly as time elapses.

Overexpression of calbindin-D28K in hippocampal progenitor cells increases neuronal differentiation and neurite outgrowth.

Excitatory stimuli are known to be a potent regulator for induction of neuronal differentiation. Calbindin-D28K buffers intracellular Ca2+ and modifies synaptic functions in neurons. However, the effects of calbindin-D28K on the regulation of activity-induced neuronal differentiation and related biochemical modifications remain unsolved. In the present study, by a gain-of-function study with retroviral vector system and dicer-generated small interfering RNA (d-siRNA) to effectively knock down the expression of calbindin-D28K, we demonstrated that calbindin-D28K at a physiologically relevant level promoted neuronal differentiation and neurite outgrowth. Increase of neuronal differentiation by calbindin-D28K overexpression was concurrent with the expression of basic helix-loop-helix (bHLH) transcriptional factors, phosphorylation of calcium and calmodulin-dependent protein kinase II (CaMKII) and NeuroD at Ser(336). KN-62, a highly specific CaMKII inhibitor, blocked the up-regulation of proneural bHLH genes, p-CaMKII, and pSer(336)NeuroD. Calbindin-D28K appeared to facilitate neuronal differentiation of both fetal and adult hippocampal progenitor cells. Together, these findings establish the novel calbindin-regulated function of CaMKII and NeuroD in control of neuronal differentiation and neurite outgrowth.

Coexistence of ulcerative colitis and Sjögren's syndrome in a patient with Takayasu's arteritis and Hashimoto's thyroiditis.

A 31-year-old woman with a 15-year history of Takayasu's arteritis (TA) and a 13-year history of Hashimoto's thyroiditis presented with hematochezia. She received a diagnosis of Sjögren's syndrome at 1 month before her visit to Kyungpook National University Medical Center. Her colonoscopic findings were compatible with a diagnosis of ulcerative colitis (UC). She was treated with oral mesalazine, and her hematochezia symptoms subsequently disappeared. The coexistence of UC and TA has been reported; however, reports on the coexistence of UC and Sjögren's syndrome, or of UC and Hashimoto's thyroiditis are rare. Although the precise etiologies of these diseases are unknown, their presence together suggests that they may have a common pathophysiologic background. Furthermore, in patients with autoimmune or vascular diseases, including TA, systemic manifestations should be assessed with consideration of inflammatory bowel diseases including UC in the presence of gastrointestinal symptoms such as diarrhea and hematochezia.

Early Activation of Phosphatidylinositol 3-Kinase after Ischemic Stroke Reduces Infarct Volume and Improves Long-Term Behavior.

Phosphatidylinositol 3-kinases (PI3Ks) have recently been implicated in apoptosis and ischemic cell death. We tested the efficacy of early intervention with a peptide PI3K activator in focal cerebral ischemia. After determining the most effective dose (24 µg/kg) and time window (2 h after MCAO) of treatment, a total of 48 rats were subjected to middle cerebral artery occlusion (MCAO). Diffusion weighted MRI (DWI) was performed 1 h after MCAO and rats with lesion sizes within a predetermined range were randomized to either PI3K activator or vehicle treatment arms. Fluid attenuated inversion recovery (FLAIR) MRI, neurological function, western blots, and immunohistochemistry were blindly assessed. Initial DWI lesion volumes were nearly identical between two groups prior to treatment. However, FLAIR showed significantly smaller infarct volumes in the PI3K activator group compared with vehicle (146 ± 81 mm3 and 211 ± 96 mm3, p = 0.045) at 48 h. The PI3K activator group also had better neurological function for up to 2 weeks. In addition, PI3K activator decreased the number of TUNEL-positive cells in the peri-infarct region compared with the control group. Western blot and immunohistochemistry showed increased expression of phosphorylated Akt (Ser473) and GSK-3ß (Ser9) and decreased expression of cleaved caspase-9 and caspase-3. Our results suggest a neuroprotective role of early activation of PI3K in ischemic stroke. The use of DWI in the randomization of experimental groups may reduce bias.

Neonatal diffusion tensor brain imaging predicts later motor outcome in preterm neonates with white matter abnormalities.

BACKGROUND: White matter (WM) abnormalities associated with prematurity are one of the most important causes of neurological disability that involves spastic motor deficits in preterm newborns. This study aimed to evaluate regional microstructural changes in diffusion tensor imaging (DTI) associated with WM abnormalities. METHODS: We prospectively studied extremely low birth weight (ELBW; <1000 g) preterm infants who were admitted to the Neonatal Intensive Care Unit of Hanyang University Hospital between February 2011 and February 2014. WM abnormalities were assessed with conventional magnetic resonance (MR) imaging and DTI near term-equivalent age before discharge. Region-of-interests (ROIs) measurements were performed to examine the regional distribution of fractional anisotropy (FA) values. RESULTS: Thirty-two out of 72 ELBW infants underwent conventional MR imaging and DTI at term-equivalent age. Ten of these infants developed WM abnormalities associated with prematurity. Five of ten of those with WM abnormalities developed cerebral palsy (CP). DTI in the WM abnormalities with CP showed a significant reduction of mean FA in the genu of the corpus callosum (p = 0.022), the ipsilateral posterior limb of the internal capsule (p = 0.019), and the ipsilateral centrum semiovale (p = 0.012) compared to normal WM and WM abnormalities without CP. In infants having WM abnormalities with CP, early FA values in neonatal DTI revealed abnormalities of the WM regions prior to the manifestation of hemiparesis. CONCLUSIONS: DTI performed at term equivalent age shows different FA values in WM regions among infants with or without WM abnormalities associated with prematurity and/or CP. Low FA values of ROIs in DTI are related with later development of spastic CP in preterm infants with WM abnormalities.

Directly converted patient-specific induced neurons mirror the neuropathology of FUS with disrupted nuclear localization in amyotrophic lateral sclerosis.

BACKGROUND: Mutations in the fused in sarcoma (FUS) gene have been linked to amyotrophic lateral sclerosis (ALS). ALS patients with FUS mutations exhibit neuronal cytoplasmic mislocalization of the mutant FUS protein. ALS patients' fibroblasts or induced pluripotent stem cell (iPSC)-derived neurons have been developed as models for understanding ALS-associated FUS (ALS-FUS) pathology; however, pathological neuronal signatures are not sufficiently present in the fibroblasts of patients, whereas the generation of iPSC-derived neurons from ALS patients requires relatively intricate procedures. RESULTS: Here, we report the generation of disease-specific induced neurons (iNeurons) from the fibroblasts of patients who carry three different FUS mutations that were recently identified by direct sequencing and multi-gene panel analysis. The mutations are located at the C-terminal nuclear localization signal (NLS) region of the protein (p.G504Wfs*12, p.R495*, p.Q519E): two de novo mutations in sporadic ALS and one in familial ALS case. Aberrant cytoplasmic mislocalization with nuclear clearance was detected in all patient-derived iNeurons, and oxidative stress further induced the accumulation of cytoplasmic FUS in cytoplasmic granules, thereby recapitulating neuronal pathological features identified in mutant FUS (p.G504Wfs*12)-autopsied ALS patient. Importantly, such FUS pathological hallmarks of the patient with the p.Q519E mutation were only detected in patient-derived iNeurons, which contrasts to predominant FUS (p.Q519E) in the nucleus of both the transfected cells and patient-derived fibroblasts. CONCLUSIONS: Thus, iNeurons may provide a more reliable model for investigating FUS mutations with disrupted NLS for understanding FUS-associated proteinopathies in ALS.