The Oncological Effect of Mutant p53 on the Metastatic Phenotype of Gastric Cancer Cells.

BACKGROUND/AIM: P53 is the most frequently mutated tumor suppressor gene among all cancers. In human cancers, specific residues of p53 are mutated at a high frequency, and those mutations are known as hotspot mutations. Mutant p53 promotes tumor progression through the gain-of-function (GOF) mechanism. However, its biological characteristics, especially its metastatic potential, owing to different hotspot mutations in gastric cancer remain unclear. In the present study, we investigated the p53-depended metastatic phenotype. MATERIALS AND METHODS: This study examined the differences in the metastatic potential of wild-type, mutant-p53-R175H, and mutant-p53-R273H NUGC-4 gastric cancer cells in vitro and in vivo. RESULTS: NUGC-4-mutant-p53-R175H cells showed significant cell proliferation, healing and invasive abilities in proliferation, wound healing and invasion assay, respectively, compared to wild-type and mutant-p53-R273H cells. Both NUGC-4-mutant-p53 cell types expressed epithelial-mesenchymal transition (EMT)-related proteins. Furthermore, NUGC-4-mutant-p53-R175H cells showed less attachment to the extracellular matrix and greater expression of EMT-related proteins than NUGC-4-mutant-p53-R273H cells. Regarding the peritoneal dissemination model, NUCG-4-mutant-p53-R175H and NUCG-4-mutant-p53-R273H cells demonstrated less frequent formation of dissemination nodules than NUGC-4-empty cells. In contrast, liver metastases were more frequent and greater in number in NUCG3-mutant-p53-R175H than in the other cell lines. CONCLUSION: Our results suggest that differences in the p53 status, even in the hotspot mutation site, affect not only the characteristics of the cells but also the metastatic ability of gastric cancer.

Agent-based approach for elucidating the release from collective arrest of cell motion in corneal epithelial cell sheet.

Changes in cell fluidity have been observed in various cellular tissues and are strongly linked to biological phenomena such as self-organization. Recent studies suggested variety of mechanisms and factors, which are still being investigated. This study aimed to investigate changes in cell fluidity in multi-layered cell sheets, by exploring the collective arrest of cell motion and its release in cultures of corneal epithelial cells. We constructed mathematical models to simulate the behaviors of individual cells, including cell differentiation and time-dependent changes in cell-cell connections, which are defined by stochastic or kinetic rules. Changes in cell fluidity and cell sheet structures were expressed by simulating autonomous cell behaviors and interactions in tissues using an agent-based model. A single-cell level spatiotemporal analysis of cell state transition between migratable and non-migratable states revealed that the release from collective arrest of cell motion was initially triggered by a decreased ability to form cell-cell connections in the suprabasal layers, and was propagated by chain migration. Notably, the disruption of cell-cell connections and stratification occurred in the region of migratable state cells. Hence, a modeling approach that considers time-dependent changes in cell properties and behavior, and spatiotemporal analysis at the single-cell level can effectively delineate emergent phenomena arising from the complex interplay of cells.

Barium-induced toxic anterior segment syndrome.

Toxic anterior segment syndrome (TASS) is a rapid-onset inflammation of the eye following uneventful ocular surgery. We report a case of TASS following Baerveldt glaucoma implant (BGI) surgery. Inductively coupled plasma-mass spectrometry (ICP-MS) identified barium in the eye and in the eluate from the bleb of the BGI. We attribute TASS in our patient to the dissolution of barium from the BGI and its entry into the eye, where it causes severe inflammation.

Time to enhancement of breast lesions and normal breast parenchyma in light of menopausal status and menstrual cycle for ultrafast dynamic contrast-enhanced MRI using compressed sensing.

PURPOSE: To assess the dependency of the Time to enhancement (TTE) of breast lesions and normal breast parenchyma from menopausal status and menstrual cycle using ultrafast compressed sensing (CS) -accelerated dynamic contrast-enhanced (DCE) MRI. METHODS: This institutional review board approved retrospective study included 89 breast cancers, 22 benign lesions and 131 normal breast parenchymal foci. A prototypical ultrafast DCE sequence obtained 30 phases with 2.9 s temporal resolution. Mean and median TTE of all breast cancers, benign lesions and normal breast parenchymal foci were assessed. we also assessed whether there were any differences in TTE regarding the menopausal status and menstrual cycle. RESULTS: The TTE of breast cancer was significantly shorter than that of benign lesions and normal breast parenchymal foci in both the premenopausal status (5.8 vs. 8.7 and 8.7 s, respectively) (p = 0.0028 and < 0.0001, respectively) and postmenopausal status (5.8 vs. 11.6 and 11.6 s, respectively) (p < 0.0001 in both). The TTE of parenchymal foci in the premenopausal status was significantly shorter than that in the postmenopausal status (p = 0.0025). Although the TTE interval between cancer and parenchymal foci in premenopausal status is shorter than that in postmenopausal status, the AUCs in the pre- and postmenopausal status for differentiating breast cancer and parenchymal foci were comparable with using different cutoff TTE values. There were no differences in TTE regarding the menstrual cycle. CONCLUSIONS: The TTE derived from ultrafast CS-accelerated DCE MRI was useful to differentiate breast cancer from benign lesions and normal breast parenchymal foci in both pre- and postmenopausal status.

Overlapping Signs and Symptoms Between Recurrent Varicella and Pityriasis rosea Gibert.

BACKGROUND: Pityriasis rosea Gibert (PRG) has features similar to those of common infectious childhood diseases, suggesting a viral cause, but no agent has been identified to date. We describe 4 children with PRG and 2 with recurrent varicella who were studied using photochronography, virology and immunology. METHODS: The 6 patients with skin rashes visited our pediatric clinic from April 2012 to May 2016. Photographs of their skin lesions were taken; blood, skin lesions, and/or nasal lavage samples were collected to detect varicella-zoster virus (VZV) DNA and antibodies; and skin tests were carried out to measure cell-mediated immunity to VZV. RESULTS: Herald patches were confirmed in 2 of 4 PRG patients. No specimen cultures were positive for infectious VZV. However, VZV-DNA was detected in skin lesions of 3 PRG patients. During the acute phase, 5 patients had IgG antibodies to VZV, and skin-test reactions were positive in 5 patients. CONCLUSIONS: IgG antibody titers to VZV at rash onset were high, suggesting that they were already rising at the appearance of the rash and that reinfection with VZV must have occurred during the prodromal stage or several weeks before rash appearance in PRG patients whose immunity had declined below the threshold. Our study suggests a new pathogenesis of PRG that might help to address incongruities of past theories on PRG sites of viral entry and replication, incubation period and variations in the clinical course of PRG from prodrome to healing.

Maximum slope of ultrafast dynamic contrast-enhanced MRI of the breast: Comparisons with prognostic factors of breast cancer.

PURPOSE: To determine the relationship between the maximum slope (MS) of ultrafast dynamic contrast-enhanced (DCE)-MRI and prognostic factors of breast cancer. METHODS: One hundred thirteen patients with 118 breast cancers were included in this study. The ultrafast DCE sequence was acquired using a higher parallel imaging factor. Its spatial resolution was 0.9 × 0.9 × 2.5 mm and its temporal resolution was 8.3 s/phase. Each lesion was automatically segmented, and the ROI of highest enhancement in the lesion was identified. In this ROI, the MS was calculated. The MS of each lesion was compared with various prognostic factors of breast cancer. RESULTS: The MS of invasive cancer (median: 9.81%/sec) was significantly higher than that of ductal carcinoma in situ (median: 7.26%/sec) (p = 0.001). In the ROC analysis, the area under the ROC curve (AUC) was 0.7295. The MS of invasive cancer with axillary lymph node (LN) metastasis (median: 11.97%/sec) was significantly higher than that without axillary LN metastasis (median: 9.425%/sec) (p = 0.0024). In the ROC analysis, the AUC was 0.7177. In addition, the MS became significantly higher as the level of the proliferation marker ki-67 increased (correlation coefficient: 0.3317) (p = 0.0009). CONCLUSIONS: MS of ultrafast DCE-MRI is useful for predicting the prognostic factors of breast cancer. Higher maximum slope (MS) is significantly associated with an invasive breast cancer component. Higher MS is significantly associated with an axillary lymph node metastasis. MS becomes significantly higher with increasing ki-67 (a proliferation marker). Ultrafast MRI is useful for predicting the prognostic factors of breast cancer.

The generation of fluorometholone nanocrystal eye drops, their metabolization to dihydrofluorometholone and penetration into rabbit eyes.

Fluorometholone is a widely used anti-inflammatory ophthalmic formulation, which elicits a lower ocular hypertensive response than other glucocorticoid medications. This serves to mitigate against the risk of steroid-induced glaucoma. Based on the hypothesis that an improved corneal permeability can increase the bioavailability of a drug, we sought to obtain fluorometholone in suspension with a small particle size. Accordingly, we describe the formulation of fluorometholone nanocrystal eye drops, which have a mean particle size of 201.2 ± 14.1 nm (standard deviation (s.d.)) when measured by dynamic light scattering. Scanning electron microscopy further indicates that fluorometholone nanocrystals are predominantly rectangular in shape. Fluorometholone microcrystals, on the other hand, with a mean particle size of 9.24 ± 4.51 µm (s.d.), tend to have a rod-like morphology. Powder x-ray diffraction revealed that fluorometholone microcrystal and nanocrystal formulations have the same crystal structure, with the main diffraction peaks at 2θ = 10.4 and 15.3°. The nanocrystal formulation was found to be stable, long-term, when stored at 10 °C for up to 6-months. High pressure liquid chromatography (HPLC) of the aqueous humor of rabbit eyes 15-240 mins after the in vivo application of fluorometholone eye drops to the ocular surface revealed that the molecule had been converted to 20α-dihydrofluorometholone (with no evidence of a 20ß-dihydrofluorometholone fraction), and that penetration was 2-6 fold higher and longer lasting with the nanocrystal, rather than the microcrystal, formulation. In current study we show how newly generated fluorometholone nanocrystals when administered as eye drops enter the anterior chamber of the eye and become metabolized to dihydrofluorometholone.

Comparison of semicircular and bent tips regarding regional differences in oscillation amplitude under various torsional power settings.

PURPOSE: To analyze the difference between the behavior of semicircular (balanced) and bent (mini) tips at 20 incremental torsional power settings. SETTING: Tsukazaki Hospital, Himeji, Japan. DESIGN: Experimental study. METHODS: Using an ultra-high-speed video camera HPV-X2, the 2 tips during torsional oscillation were recorded, comparing tip behavior at power settings from 5% to 100% by tracking points 1 to 5 (tip end and at 1325, 2650, 3975, and 5035 µm from the tip end). RESULTS: Both tips increased their amplitude widths, drawing an S-curve at all points as the torsional power setting was increased, reaching their upper limits from 70% to 90% torsional power. At all 20 power settings, both tips showed significantly different amplitudes (all P < .01), and the difference of the amplitude increased as the power setting increased. Although, at points 1 and 3, the balanced tip amplitude was nearly 1.5 times larger than the mini tip amplitude, the amplitude difference was 10 µm or less at points 2 and 4. At point 5, the mini tip amplitude was at least 3 times more than the balanced tip amplitude. CONCLUSIONS: The amplitude does not increase proportionally and varies markedly with the tip shape on reaching the upper limit, suggesting that a higher power setting might not contribute greatly to nuclear fragmentation. The balanced tip might cause greater damage to surrounding tissues if it is inserted at approximately 3 mm from the wound site. To obtain maximum shaft stability using the balanced tip, it is important to insert at least 5 mm.

Cell jamming, stratification and p63 expression in cultivated human corneal epithelial cell sheets.

Corneal limbal epithelial stem cell transplantation using cultivated human corneal epithelial cell sheets has been used successfully to treat limbal stem cell deficiencies. Here we report an investigation into the quality of cultivated human corneal epithelial cell sheets using time-lapse imaging of the cell culture process every 20 minutes over 14 days to ascertain the level of cell jamming, a phenomenon in which cells become smaller, more rounded and less actively expansive. In parallel, we also assessed the expression of p63, an important corneal epithelial stem cell marker. The occurrence of cell jamming was variable and transient, but was invariably associated with a thickening and stratification of the cell sheet. p63 was present in all expanding cell sheets in the first 9 days of culture, but it's presence did not always correlate with stratification of the cell sheet. Nor did p63 expression necessarily persist in stratified cell sheets. An assessment of cell jamming, therefore, can shed significant light on the quality and regenerative potential of cultivated human corneal epithelial cell sheets.

The secretome of adipose-derived mesenchymal stem cells attenuates epithelial-mesenchymal transition in human corneal epithelium.

INTRODUCTION: Epithelial-mesenchymal transition (EMT) induces the loss of cell-cell interactions in polarized epithelial cells and converts these cells to invasive mesenchymal-like cells. It is also involved in tissue fibrosis including that occurring in some ocular surface diseases such as pterygium and in subepithelial corneal fibrosis in limbal stem cell deficiency. Here, we examined the effects of the secretome of human adipose-derived mesenchymal stem cells (AdMSCs) on EMT in human corneal epithelial cells (CECs). METHODS: EMT was induced with transforming growth factor-ß (TGF-ß) in primary human CECs isolated from the human corneal limbus. The effects of the AdMSC secretome on EMT in these cells or stratified CEC sheets were analyzed by co-cultivation experiments with the addition of AdMSC conditioned-medium. The expression of EMT-related genes and proteins in CECs was analyzed. The superstructure of CECs was observed by scanning electron microscopy. Furthermore, the barrier function of CEC sheets was analyzed by measuring transepithelial electrical resistance (TER). RESULTS: The AdMSC secretome was found to suppress EMT-related gene expression and attenuate TGF-ß-induced corneal epithelial dysfunction including the dissociation of cell-cell interactions and decreases in TER in constructed CEC sheets. CONCLUSIONS: The secretome of AdMSCs can inhibit TGF-ß-induced EMT in CECs. These findings suggest that this could be a useful source for the treatment for EMT-related ocular surface diseases.

Adverse Effects on the Postoperative Urinary Function After Combined Resection of Inferior Vesical Artery in Laparoscopic Lateral Pelvic Lymph Node Dissection: Retrospective Analysis of Consecutive 95 Series.

BACKGROUND: The combined resection of the vesical artery (VA) in laparoscopic lateral pelvic lymph node dissection (L-LPLD) was reported to facilitate the safe dissection of metastatic lymph nodes. However, whether or not the combined VA resection affects the urinary function remains controversial. PURPOSE: The purpose of the present study was to examine the risk factors for the postoperative urinary dysfunction (PUD) after L-LPLD followed by total mesorectal excision and to clarify the effects of the combined VA resection in L-LPLD on PUD. PATIENTS AND METHODS: L-LPLD was performed in 95 patients with advanced rectal cancer at Saga University Hospital and Kyushu University Hospital from January 2013 to December 2017. The risk factors for PUD after L-LPLD were investigated. RESULTS: The univariate analysis revealed that the combined resection of the inferior vesical artery (IVA) was a risk factor for PUD. To examine by the type of IVA resection, the incidence of PUD significantly increased with the bilateral IVA resection, but the unilateral IVA resection induced PUD on the same level with the preservation of IVA. CONCLUSIONS: Bilateral IVA resection in L-LPLD could increase the incidence of PUD. Thus, if possible, the preservation of the unilateral IVA through L-LPLD should be considered.

Is additional mesial temporal resection necessary for intractable epilepsy with cavernous malformations in the temporal neocortex?

PURPOSE: Cavernous malformation (CM) in the temporal neocortex causes intractable epilepsy. Whether to resect additional mesial temporal structures in addition to the lesionectomy is a still controversial issue. To clarify the need for the procedure, we retrospectively analyzed pre- and postoperative clinical data of patients with surgically removed CM. SUBJECTS AND METHODS: We included data from 18 patients with CM in the temporal neocortex who presented with intractable epilepsy. Eleven patients of our early series were treated with extended resection, i.e., lesionectomy and the resection of additional mesial temporal structures. Seven patients underwent lesionectomy, i.e., removal of the CM and of hemosiderin-stained surrounding brain tissue. Pathological assessments of the resected hippocampus were performed. Chronic intracranial electroencephalography (EEG) recordings were obtained in 6 patients. We performed perioperative neuropsychological assessments in all patients. RESULTS: The seizure outcome was recorded as Engel class I in 17 patients (94.4%); Ia = 12 (66.7%) Ib = 2 (11.1%), Ic = 1 (5.6%), Id = 2 (11.1%), and class IIb in one patient (5.6%). Adding resection of the mesial temporal structures to lesionectomy did not alter the seizure outcome. Pathology of hippocampus revealed limited neuronal loss in CA4. Ictal onsets in the ipsilateral lateral cortex were detected in all 6 patients who underwent intracranial EEG. In 4 patients each, we also detected ictal onsets from the ipsilateral mesial temporal structures and from the contralateral temporal lobe. Postoperatively, in the patients where their CM was located in the language-dominant hemisphere (n = 10), the full-scale intelligence quotient (IQ) and the performance IQ increased (p < 0.05), whereas the verbal memory (WMS-R) deteriorated in two of 5 patients. CONCLUSION: Excellent seizure outcomes were obtained even the lesionectomy alone. To confirm appropriate surgical strategy for lateral temporal CM with intractable epilepsy, further studies in large sample size are needed.

Risk factors of cognitive impairment in pediatric epilepsy patients with focal cortical dysplasia.

OBJECTIVE: The purpose of this study was to identify the risk factors of cognitive impairment in pediatric epilepsy patients with focal cortical dysplasia (FCD). METHODS: 77 patients with histopathologically confirmed FCD were studied. The statistical relationship between cognition levels and clinical factors at presurgical evaluation was analyzed. Cognitive function was evaluated by development quotient or intelligence quotient (DQ-IQ). RESULTS: Ages at seizure onset were younger than 15 years (mean ±â€¯SD; 5.0 ±â€¯4.2 years). Mean disease duration was 14.5 ±â€¯8.5 years. Mean age at pre-surgical DQ-IQ evaluation was 34.8 ±â€¯10.7 years. Mean DQ-IQ was 60.5 ±â€¯20.5, and 41 of 77 (53.2%) patients had mental retardation (DQ-IQ < 70). Younger seizure onset and seizure clustering were significantly associated with lower DQ-IQ (p < 0.001). A multiple regression study identified higher seizure frequency pattern, a history of epileptic spasm and status epilepticus as aggravating factors of DQ-IQ decline (R2 = 0.63, p < 0.001). On the other hand, the risk was decreased in patients with habitual focal aware seizure and transient seizure-free periods up to 6 months in the course of epilepsy. FCD location (FCD site, extent of radiological lesion and laterality) and histopathology of FCD did not affect DQ-IQ. CONCLUSIONS: Our study suggests that seizure characteristics including higher seizure frequency pattern, a history of epileptic spasm, status epilepticus, seizure clustering and early onset of seizure are risk factors of cognitive impairment in FCD patients.

Andrographolide induces degradation of mutant p53 via activation of Hsp70.

The tumor suppressor gene p53 encodes a transcription factor that regulates various cellular functions, including DNA repair, apoptosis and cell cycle progression. Approximately half of all human cancers carry mutations in p53 that lead to loss of tumor suppressor function or gain of functions that promote the cancer phenotype. Thus, targeting mutant p53 as an anticancer therapy has attracted considerable attention. In the current study, a small-molecule screen identified andrographlide (ANDRO) as a mutant p53 suppressor. The effects of ANDRO, a small molecule isolated from the Chinese herb Andrographis paniculata, on tumor cells carrying wild-type or mutant p53 were examined. ANDRO suppressed expression of mutant p53, induced expression of the cyclin-dependent kinase inhibitor p21 and pro-apoptotic proteins genes, and inhibited the growth of cancer cells harboring mutant p53. ANDRO also induced expression of the heat-shock protein (Hsp70) and increased binding between Hsp70 and mutant p53 protein, thus promoting proteasomal degradation of p53. These results provide novel insights into the mechanisms regulating the function of mutant p53 and suggest that activation of Hsp70 may be a new strategy for the treatment of cancers harboring mutant p53.

Successful hemispherotomy in two refractory epilepsy patients with cerebral hemiatrophy and contralateral EEG abnormalities.

We describe two cases of refractory epilepsy with cerebral hemiatrophy and contralateral electroencephalographic (EEG) abnormalities, in which hemispherotomy of the atrophic hemisphere effectively controlled seizures. Case 1 was a 5-year-1-month-old girl with refractory bilateral asymmetrical tonic posturing seizures predominantly in the right arm. Magnetic resonance imaging showed left porencephaly corresponding to a left middle cerebral artery infarction. Case 2 was a 3-year-8-month-old boy with refractory bilateral asymmetrical tonic posturing seizures predominantly in the right arm due to atrophy of the left cerebral hemisphere after septic meningitis. Both patients had right hemiparesis and was incapable of pinching by the right hand. Contralateral interictal and ictal EEG abnormalities were observed. Interictal 99mTc-ethyl cysteinate dimer (99mTc-ECD) single photon emission computed tomography (SPECT) showed hypoperfusion and ictal 99mTc-ECD-SPECT showed hyperperfusion within the left cerebral hemisphere. Left hemispherotomy was performed. Cases 1 and 2 remained seizure-free at the last follow-up 18 months and 15 months, respectively, after surgery, and contralateral interictal EEG abnormalities disappeared. In patients with cerebral hemiatrophy and contralateral EEG abnormalities, epilepsy surgery may be considered when the laterality of seizure semiology, functional imaging findings and motor deficits were concordant with the atrophic side. Ictal SPECT is effective to confirm the epileptogenic hemisphere.

Hypoxia-induced ANGPTL4 sustains tumour growth and anoikis resistance through different mechanisms in scirrhous gastric cancer cell lines.

Patients with scirrhous gastric cancer (SGC) frequently develop peritoneal dissemination, which leads to poor prognosis. The secreted protein angiopoietin-like-4 (ANGPTL4), which is induced by hypoxia, exerts diverse effects on cancer progression. Here, we aimed to determine the biological function of ANGPTL4 in SGC cells under hypoxia. ANGPTL4 levels were higher in SGC cells under hypoxia than in other types of gastric cancer cells. Hypoxia-induced ANGPTL4 mRNA expression was regulated by hypoxia-inducible factor-1α (HIF-1α). Under hypoxic conditions, monolayer cultures of ANGPTL4 knockdown (KD) 58As9 SGC (58As9-KD) cells were arrested in the G1 phase of the cell cycle through downregulation of c-Myc and upregulation of p27, in contrast to control 58As9-SC cells. Moreover, the ability of 58As9-KD xenografts to form tumours in nude mice was strongly suppressed. When 58As9-KD cells were cultured in suspension, hypoxia strongly increased their susceptibility to anoikis through suppression of the FAK/Src/PI3K-Akt/ERK pro-survival pathway, followed by activation of the apoptotic factors caspases-3, -8 and -9. The development of peritoneal dissemination by 58As9-KD cells was completely inhibited compared with that by 58As9-SC cells. In conclusion, ANGPTL4 is uniquely induced by hypoxia in cultured SGC cells and is essential for tumour growth and resistance to anoikis through different mechanisms.

A Self-Assembling Peptide Gel as a Vitreous Substitute: A Rabbit Study.

Purpose: To evaluate a self-assembling peptide gel as a potential vitreous substitute. Methods: PanaceaGel SPG-178, a self-assembling peptide gel, was diluted with distilled water and a balanced salt solution to achieve a final peptide concentration of 0.1%. The gel's refractive index, visible light transmission rate, and rheologic properties were investigated. The gel's biocompatibility was evaluated by examining the cellular viability (live and dead staining) and proliferation rate (alamarBlue assay). A 25-G pars plana vitrectomy was performed on the right eye of 21 New Zealand white rabbits. The gel was then injected into the vitreous cavity of 15 eyes. Six eyes were injected with a balanced salt solution (BSS) and served as controls. Toxicity was examined using electroretinography and histologic analysis after the injection of the gel. Results: The gel's physical properties closely resembled those of human vitreous. The gel showed no apparent toxicity. When the gel was injected into the vitreous cavity, fragmentation was not observed. Additionally, the gel remained transparent in the vitreous cavity and no complications were observed for 3 months after the injection. Electroretinography and histology confirmed the gel's biocompatibility. Conclusions: This diluted self-assembling peptide gel could be provide a promising vitreous substitute.

Long-term outcomes of epilepsy surgery in 85 pediatric patients followed up for over 10 years: a retrospective survey.

OBJECTIVE The aim of this study was to investigate the treatment outcomes and social engagement of patients who had undergone pediatric epilepsy surgery more than 10 years earlier. METHODS Between 1983 and 2005, 110 patients younger than 16 years underwent epilepsy surgery at the National Epilepsy Center, Shizuoka Institute of Epilepsy and Neurological Disorders. The authors sent a questionnaire to 103 patients who had undergone follow-up for more than 10 years after surgery; 85 patients (82.5%) responded. The survey contained 4 categories: seizure outcome, use of antiepileptic drugs, social participation, and general satisfaction with the surgical treatment (resection of the epileptic focus, including 4 hemispherectomies). The mean patient age at the time of surgery was 9.8 ± 4.2 (SD) years, and the mean duration of postoperative follow-up was 15.4 ± 5.0 years. Of the 85 patients, 79 (92.9%) presented with a lesional pathology, such as medial temporal sclerosis, developmental/neoplastic lesions, focal cortical dysplasia, and gliosis in a single lobe. RESULTS For 65 of the 85 responders (76.5%), the outcome was recorded as Engel Class I (including 15 [93.8%] of 16 patients with medial temporal sclerosis, 20 [80.0%] of 25 with developmental/neoplastic lesions, and 27 [73.0%] of 37 with focal cortical dysplasia). Of these, 29 (44.6%) were not taking antiepileptic drugs at the time of our survey, 29 (44.6%) held full-time jobs, and 33 of 59 patients (55.9%) eligible to drive had a driver's license. Among 73 patients who reported their degree of satisfaction, 58 (79.5%) were very satisfied with the treatment outcome. CONCLUSIONS The seizure outcome in patients who underwent resective surgery in childhood and underwent followup for more than 10 years was good. Of 85 respondents, 65 (76.5%) were classified in Engel Class I. The degree of social engagement was relatively high, and the satisfaction level with the treatment outcome was also high. From the perspective of seizure control and social adaptation, resective surgery yielded longitudinal benefits in children with intractable epilepsy, especially those with a lesional pathology in a single lobe.

Capturing and concentrating adenovirus using magnetic anionic nanobeads.

We recently demonstrated how various enveloped viruses can be efficiently concentrated using magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic anhydrate). However, the exact mechanism of interaction between the virus particles and anionic beads remains unclear. To further investigate whether these magnetic anionic beads specifically bind to the viral envelope, we examined their potential interaction with a nonenveloped virus (adenovirus). The beads were incubated with either adenovirus-infected cell culture medium or nasal aspirates from adenovirus-infected individuals and then separated from the supernatant by applying a magnetic field. After thoroughly washing the beads, adsorption of adenovirus was confirmed by a variety of techniques, including immunochromatography, polymerase chain reaction, Western blotting, and cell culture infection assays. These detection methods positively identified the hexon and penton capsid proteins of adenovirus along with the viral genome on the magnetic beads. Furthermore, various types of adenovirus including Types 5, 6, 11, 19, and 41 were captured using the magnetic bead procedure. Our bead capture method was also found to increase the sensitivity of viral detection. Adenovirus below the detectable limit for immunochromatography was efficiently concentrated using the magnetic bead procedure, allowing the virus to be successfully detected using this methodology. Moreover, these findings clearly demonstrate that a viral envelope is not required for binding to the anionic magnetic beads. Taken together, our results show that this capture procedure increases the sensitivity of detection of adenovirus and would, therefore, be a valuable tool for analyzing both clinical and experimental samples.

Impaired mitophagy activates mtROS/HIF-1α interplay and increases cancer aggressiveness in gastric cancer cells under hypoxia.

Mitochondrial autophagy (mitophagy) is a selective form of autophagy and a critical step in excluding mitochondria damaged by stress, including hypoxia. This study aimed to determine whether the integrity of mitophagy affected production of the mitochondrial reactive oxygen species (mtROS), hypoxia inducible factor (HIF)-1α expression and aggressive characteristics in GC cells under hypoxia. Three GC cell lines, 44As3, 58As9 and MKN45, were investigated in this study. HIF-1α expression was induced in the three GC cell lines under hypoxia, with higher expression observed in 44As3 and 58As9 cells compared with MKN45 cells. Cell survival and invasion abilities under hypoxia were significantly stronger in 44As3 and 58As9 cells than MKN45 cells. Moreover, mtROS accumulated in a time-dependent manner in 44As3 and 58As9 cells, but not in MKN45 cells. ROS scavenger N-acetyl-L-cysteine (NAC) treatment resulted in strong attenuation of HIF-1α expression, whereas HIF-1α knockdown increased ROS production in the three GC cell lines under hypoxia. These results suggested that the mtROS/HIF-1α interplay affected the hypoxia-induced cancer aggressiveness. Assessment of mitophagy by LC3-I/II conversion, SQSTM1/p62 degradation and specific fluorescence markers demonstrated that hypoxia-induced mitophagy was observed only in MKN45 cells, while the process was impaired in the other two cell lines. Treatment with the autophagy inhibitor chloroquine conversely increased HIF-1α expression, mtROS generation, cell survival and invasion in hypoxic MKN45 cells. The present study revealed a novel mechanism in which the integrity of mitophagy might determine cancer aggressiveness via mtROS/HIF-1α interplay in GC cells under hypoxic conditions.