Dynamic Imaging of Lipid Droplets in Cells and Tissues by Using Dioxaborine Barbiturate-Based Fluorogenic Probes.

Lipids are essential for various cellular functions, including energy storage, membrane flexibility, and signaling molecule production. Maintaining proper lipid levels is important to prevent health problems such as cancer, neurodegenerative disorders, cardiovascular diseases, obesity, and diabetes. Monitoring cellular lipid droplets (LDs) in real-time with high resolution can provide insights into LD-related pathways and diseases owing to the dynamic nature of LDs. Fluorescence-based imaging is widely used for tracking LDs in live cells and animal models. However, the current fluorophores have limitations such as poor photostability and high background staining. Herein, we developed a novel fluorogenic probe based on a push-pull interaction combined with aggregation-induced emission enhancement (AIEE) for dynamic imaging of LDs. Probe 1 exhibits favorable membrane permeability and spectroscopic characteristics, allowing specific imaging of cellular LDs and time-lapse imaging of LD accumulation. This probe can also be used to examine LDs in fruit fly tissues in various metabolic states, serving as a highly versatile and specific tool for dynamic LD imaging in cellular and tissue environments.

PREVALENCE AND RISK FACTORS OF AGE-RELATED MACULAR DEGENERATION FEATURES AMONG PILOTS.

PURPOSE: To investigate the prevalence and risk factors of age-related macular degeneration features among pilots of Republic of Korea Air Force. METHODS: This retrospective, cross-sectional study was performed with a total of 2781 Republic of Korea Air Force pilots who underwent regular medical examinations between 2020 and 2021. Age-related macular degeneration features were determined and graded by fundus photographs. Risk factors were identified with logistic regression analysis in odds ratio (OR) and 95% confidence interval (CI). RESULTS: The prevalence was 12.9% in the Republic of Korea Air Force pilots and 35.2% in those older than 50 years. Pilots with age-related macular degeneration features were positively associated with age (OR: 1.082, CI: 1.067-1.096, P < 0.001), male sex (OR: 0.229, CI: 0.056-0.939, P = 0.041), smoking (OR: 1.027, CI: 1.008-1.047, P = 0.006), flight time (OR: 1.004, CI: 1.003-1.005, P < 0.001), total cholesterol (OR: 1.004, CI: 1.000-1.007, P = 0.033), and low-density lipoprotein (OR: 1.005, CI: 1.001-1.008, P = 0.011). Aircraft type was also identified as a risk factor (OR: 0.617, CI: 0.460-0.827 for carrier, OR: 0.572, CI: 0.348-0.940 for helicopter, P = 0.002), with fighter pilots having a higher risk than carrier and helicopter pilots. The results were similar for pilots older than 50 years. CONCLUSION: The prevalence of age-related macular degeneration features in Republic of Korea Air Force pilots was higher than in other general populations studied. Identified risk factors such as flight time and aircraft type suggest potential occupational risk of age-related macular degeneration in aviators.

Artificial Intelligence-Based Anomaly Detection Technology over Encrypted Traffic: A Systematic Literature Review.

As cyber-attacks increase in unencrypted communication environments such as the traditional Internet, protected communication channels based on cryptographic protocols, such as transport layer security (TLS), have been introduced to the Internet. Accordingly, attackers have been carrying out cyber-attacks by hiding themselves in protected communication channels. However, the nature of channels protected by cryptographic protocols makes it difficult to distinguish between normal and malicious network traffic behaviors. This means that traditional anomaly detection models with features from packets extracted a deep packet inspection (DPI) have been neutralized. Recently, studies on anomaly detection using artificial intelligence (AI) and statistical characteristics of traffic have been proposed as an alternative. In this review, we provide a systematic review for AI-based anomaly detection techniques over encrypted traffic. We set several research questions on the review topic and collected research according to eligibility criteria. Through the screening process and quality assessment, 30 research articles were selected with high suitability to be included in the review from the collected literature. We reviewed the selected research in terms of dataset, feature extraction, feature selection, preprocessing, anomaly detection algorithm, and performance indicators. As a result of the literature review, it was confirmed that various techniques used for AI-based anomaly detection over encrypted traffic were used. Some techniques are similar to those used for AI-based anomaly detection over unencrypted traffic, but some technologies are different from those used for unencrypted traffic.

Electrochemical Detection of Dopamine Release from Living Neurons Using Graphene Oxide-Incorporated Polypyrrole/Gold Nanocluster Hybrid Nanopattern Arrays.

Stem-cell-based therapeutics have shown immense potential in treating various diseases that are currently incurable. In particular, partial recovery of Parkinson's disease, which occurs due to massive loss or abnormal functionality of dopaminergic (DAnergic) neurons, through the engraftment of stem-cell-derived neurons ex vivo is reported. However, precise assessment of the functionality and maturity of DAnergic neurons is still challenging for their enhanced clinical efficacy. Here, a novel conductive cell cultivation platform, a graphene oxide (GO)-incorporated metallic polymer nanopillar array (GOMPON), that can electrochemically detect dopamine (DA) exocytosis from living DAnergic neurons, is reported. In the cell-free configuration, the linear range is 0.5-100 µm, with a limit of detection of 33.4 nm. Owing to its excellent biocompatibility, a model DAnergic neuron (SH-SY5Y cell) can be cultivated and differentiated on the platform while their DA release can be quantitatively measured in a real-time and nondestructive manner. Finally, it is showed that the functionality of the DAnergic neurons derived from stem cells can be precisely assessed via electrochemical detection of their DA exocytosis. The developed GOMPON is highly promising for a wide range of applications, including real-time monitoring of stem cell differentiation into neuronal lineages, evaluating differentiation protocols, and finding practical stem cell therapies.

Comparison of the Incidence of Nd:YAG Laser Capsulotomy Based on the Type of Intraocular Lens.

Background and Objectives: Posterior capsular opacification (PCO) is the most common long-term complication of successful cataract surgery and can cause visual impairment. We aimed to investigate the effects of intraocular lens (IOL) characteristics on PCO by comparing the incidence of neodymium-doped yttrium aluminum garnet (Nd:YAG) laser capsulotomy for different types of intraocular lenses. Materials and Methods: A retrospective analysis was performed on 2866 eyes that underwent cataract surgery between January 2010 and December 2017, with at least 5 years of follow-up. The IOLs used for surgery were the hydrophobic lenses SN60WF (Alcon, Fort Worth, TX, USA), ZCB00 (Johnson & Johnson Vision, Santa Ana, CA, USA), and MX60 (Bausch & Lomb, Rochester, NY, USA), and the hydrophilic lens MI60 (Bausch & Lomb, Rochester, NY, USA). We analyzed the incidence of Nd:YAG laser capsulotomy according to the type of IOL used. Results: The incidence of Nd:YAG laser capsulotomy was significantly higher with MI60 lenses (31.70%, 175/552 eyes) compared to SN60WF (7.90%, 113/1431 eyes), ZCB00 (10.06%, 64/636 eyes), and MX60 (10.57%, 13/123 eyes; p < 0.001) lenses. The incidence of Nd:YAG laser capsulotomy was significantly lower with the hydrophobic IOLs (8.68%, 190/2190 eyes) than with the hydrophilic IOL (31.70%, 175/552 eyes; p < 0.001). Over time, the rate of increase in the cumulative number of Nd:YAG laser capsulotomy cases was the highest with MI60. The cumulative rate of Nd:YAG laser capsulotomy during the first 3 years was 4.90% with SN60WF (70/1431 eyes), 6.76% with ZCB00 (43/636 eyes), 8.94% with MX60 (11/123 eyes), and 26.10% with MI60 (144/552 eyes) lenses. Conclusions: The incidence of PCO is influenced by the material of the IOLs. The hydrophilic IOL was associated with a higher rate of Nd:YAG laser capsulotomy than the hydrophobic IOLs, with a shorter time to Nd:YAG laser capsulotomy.

Establishment of a useful in vitro decidual induction model using eCG-primed nonpregnant mouse endometrial stromal cells†.

Uterine endometrial differentiation is essential for developmental continuity and female health. A convenient in vitro model mimicking the physiological status is needed to effectively evaluate implantation and uterine response mechanisms. Thus, we developed a promising in vitro model, the FSS (FSH mimic-stimulated synchronized) model, by using primary mouse uterine stromal cells (mUSCs) obtained from equine chorionic gonadotropin (eCG)-primed mice. These mUSCs could be differentiated into decidualized cells with 17 beta-estradiol (E2) and progesterone (P4). The pregnancy day 4 (PD4) model, in which mUSCs are obtained at day 4 of pregnancy, was used as a control. The cell shape index and polyploidy rates were similar between the two models. The staining intensities of lipids and glycogen were significantly higher in the induced groups in both models but stronger in the FSS model than in the PD4 model. The expression levels of AP-TNAP, cathepsin L, Prl8a2, Gja1, Cebpb, and Igfbp1 were increased at 24 h after decidual induction. PR-alpha and PR-beta levels were also increased at 24 h after decidual induction in both models. These results indicate that the FSS model provides a convenient method for obtaining USCs that are usable for various experimental approaches due to their physiological competence and flexibility for triggering induction. This may serve as a model system for the study of pathogeneses originating from the endometrium or communication with other tissues and lead to a better understanding of embryo implantation mechanisms. Furthermore, the results of this study will be integral for further refinements of 3D uterine culture manipulation techniques.

Familial Risk of Inflammatory Bowel Disease: A Population-Based Cohort Study in South Korea.

BACKGROUND & AIMS: Despite the rapid increase in inflammatory bowel disease (IBD), population-level familial risk estimates of IBDs still are lacking in Asian-Pacific countries. We aimed to quantify the familial risk of incident IBD among first-degree relatives (FDRs) of individuals with IBD according to age, sex, and familial relationship. METHODS: Using the South Korea National Health Insurance database (2002-2017), which has complete population coverage and confirmed accuracy of both FDR information and IBD diagnoses, we constructed a cohort of 21,940,795 study subjects comprising 12 million distinct families. We calculated incidence risk ratios of ulcerative colitis (UC) or Crohn's disease (CD) in individuals of affected FDRs compared with individuals without affected FDRs. RESULTS: Of 45,717 individuals with UC and 17,848 individuals with CD, 3.8% and 3.1% represented familial cases, respectively. Overall, there was a 10.2-fold (95% CI, 9.39-11.1) and a 22.1-fold (95% CI, 20.5-24.5) significantly higher adjusted risk of UC and CD among FDRs of individuals with vs without IBD. Familial risk was highest among twins, followed by nontwin siblings, and then offspring of affected parents. Familial risk generally was higher within generations (sibling-sibling) vs between generations (parent-offspring). Familial risk also increased with the increasing number of affected FDRs. CONCLUSIONS: According to this population-based analysis, there is a substantially increased risk of IBD among FDRs of affected individuals, with the highest risk among siblings and for CD. These findings might help with an earlier diagnosis and appropriate therapeutic intervention in FDRs of individuals with IBD. Dedicated studies are needed to evaluate the contributions of shared early-in-life environmental exposures and genetic factors.

Familial risk of Behçet's disease among first-degree relatives: a population-based aggregation study in Korea.

OBJECTIVE: Previous studies have indicated that Behçet's disease (BD) has a genetic component, however population-level familial risk estimates are unavailable. We quantified the familial incidence and risk of BD in first-degree relatives (FDR) according to age, sex and type of family relationship. METHODS: Using the Korean National Health Insurance database, which has full population coverage and confirmed FDR information, we constructed a cohort of 21 940 795 individuals comprising 12 million families, which were followed for a familial occurrence of BD from 2002 to 2017. Age- and sex-adjusted incidence risk ratios for BD were calculated in individuals with affected FDR compared with those without affected FDR. RESULTS: Among the total study population, 53 687 individuals had affected FDR, of whom 284 familial cases developed BD with an incidence of 3.57/104 person-years. The familial risk (incidence) for BD was increased to 13.1-fold (2.71/104 person-years) in individuals with an affected father, 13.9-fold (3.11/104 person-years) with affected mother, 15.2-fold (4.9/104 person-years) with an affected sibling and the highest risk was 165-fold (46/104 person-years) with an affected twin. Familial risks showed age dependence, being higher in younger age groups. The sex-specific familial risk was similar in males and females. CONCLUSION: This study provides quantified estimates of familial incidence and risk in FDR of BD patients in an entire population. Familial risks were higher within generation (sibling-sibling) vs between generations (parent-offspring). This implicates complex interactions between genetic factors and shared childhood environmental exposures in the pathogenesis of BD.

Incidence, mortality, and cardiovascular diseases in pituitary adenoma in Korea: a nationwide population-based study.

PURPOSE: Few nationally representative studies have evaluated the epidemiology of PA (pituitary adenoma). This South Korean study evaluated the incidence of different PA subtypes, cardiovascular disease (CVD), and related mortality. METHODS: This population-based study evaluated 31,898 patients with PA during 2005-2015. The incidence of PA, mortality, and CVD occurrence in PA cases were evaluated during a median follow-up of 5.3 years (range: 0-10 years). Cox regression analysis was used to evaluate the associations between CVD and mortality. RESULTS: The annual incidences (per 100,000 population) were 3.5 for non-functioning pituitary adenoma (NFPA), 1.6 for prolactinoma (PRL), 0.5 for growth hormone-secreting pituitary adenoma (GH), and 0.2 for adrenocorticotropic or thyroid-stimulating hormone-secreting pituitary adenoma (ACTH + TSH). The standardized mortality ratios were 1.9 for ACTH + TSH, 1.7 for NFPA with hypopituitarism, 1.4 for NFPA without hypopituitarism, 1.3 for GH, and 1.1 for PRL. During 2005-2015, the overall incidence of CVD among PA patients was 6.6% (2106 cases), and the standardized incidence ratios were 4.1 for hemorrhagic stroke, 3.0 for ischemic stroke, and 1.7 for acute myocardial infarction. The standardized incidence ratios for stroke were significantly higher in the ACTH + TSH and NFPA groups, which also had higher risks of CVD-related mortality, relative to the PRL and GH groups. CONCLUSION: South Korea had a relatively high incidence of NFPA. The incidence of stroke was highest for ACTH + TSH and NFPA, which was directly related to mortality during long-term follow-up. Patients with these types of PA should receive stroke prevention measures to reduce their risk of mortality.

EFFECTS OF GLYCEMIC VARIABILITY ON THE PROGRESSION OF DIABETIC RETINOPATHY AMONG PATIENTS WITH TYPE 2 DIABETES.

PURPOSE: To evaluate the effects of glycemic variability on the progression of diabetic retinopathy (DR) among individuals with Type 2 diabetes and to test the hypothesis that consistent glycemic control delays the progression of DR. METHODS: This retrospective study included 1,125 participants with a follow-up period of more than 5 years and more than 20 glucose laboratory test results. The hazard ratio of ≥3 steps of progression on the Early Treatment Diabetic Retinopathy Study person scale and progression to proliferative DR were assessed. RESULTS: An increase in the HbA1c SD was associated with a higher risk of ≥3 step progression (P < 0.001) and progression to proliferative DR (P < 0.001). Not only mean HbA1c, but also HbA1c SD was associated with a lower risk of ≥3 steps of progression (P < 0.001), and progression to proliferative DR (P < 0.001). CONCLUSION: Achievable and consistent glycemic control may contribute to the delay in DR progression. CLINICAL TRIAL REGISTRATION NUMBER: Institutional review board of Inje University (No. 202003014).

TOPOGRAPHIC CHANGES IN CHOROIDAL THICKNESS IN AGE-RELATED MACULAR DEGENERATION DURING THE DEVELOPMENT OF ACTIVE CHOROIDAL NEOVASCULARIZATION.

PURPOSE: To evaluate topographic changes in choroidal thickness during development of choroidal neovascularization (CNV) in treatment-naive age-related macular degeneration (AMD) and to test the value of such changes as a predictive tool of CNV development. METHODS: This retrospective cohort included 86 eyes that developed CNV from intermediate AMD, 43 eyes with intermediate AMD, and 36 eyes without AMD. Patients with intermediate AMD underwent spectral domain optical coherence tomography using enhanced depth imaging mode every 6 months until CNV was detected. Choroidal neovascularization was localized to one of the subfields of Early Treatment of Diabetic Retinopathy Study grid on fluorescein angiography. Average choroidal thickness of each subfield was calculated. RESULTS: Choroidal thickness of the subfield where CNV developed at first clinical detection significantly increased compared with that 6 months before (P = 0.000 for central, P = 0.001 for superior parafoveal, P = 0.002 for temporal parafoveal, P = 0.002 for inferior parafoveal, and P = 0.001 for nasal parafoveal subfield). In eight patients who visited unexpectedly 3 months before CNV development in central subfield, choroidal thickness of central subfield increased significantly compared with that 6 months before CNV development (P = 0.001). CONCLUSION: Choroidal neovascularization development accompanied choroidal thickening of the corresponding subfield. Regular measurement of choroidal thickness may assist in prediction of CNV.

Tissue-specific roles of GCN2 in aging and autosomal dominant retinitis pigmentosa.

The organisms have the capacity to sense and adapt to their surroundings for their life in a dynamic environment. In response to amino acid starvation, cells activate a rectifying physiological program, termed the integrated stress response (ISR), to restore cellular homeostasis. General controlled non-repressed (GCN2) kinase is a master regulator of the ISR and modulates protein synthesis in response to amino acid starvation. We previously established the GCN2/ATF4/4E-BP pathway in development and aging. Here, we investigated the tissue-specific roles of GCN2 upon dietary restriction of amino acid in a Drosophila model. The knockdown of GCN2 in the gut and fat body, an energy sensing organ in Drosophila, abolished the beneficial effect of GCN2 in lifespan extension upon dietary restriction of amino acids. Proteome analysis in an autosomal dominant retinitis pigmentosa (ADRP) model showed that dietary restriction of amino acids regulates the synthesis of proteins in several pathways, including mitochondrial translation, mitochondrial gene expression, and regulation of biological quality, and that gcn2-mutant flies have reduced levels of these mitochondria-associated proteins, which may contribute to retinal degeneration in ADRP. These results indicate that the tissue-specific regulation of GCN2 contributes to normal physiology and ADRP progression.

Accuracy of swept-source optical coherence tomography based biometry for intraocular lens power calculation: a retrospective cross-sectional study.

BACKGROUND: To evaluate the accuracy of biometric measurements by a swept-source optical coherence tomography (SS-OCT) based biometry for intraocular lens (IOL) power calculation. METHODS: This retrospective observational study enrolled 431 patients undergoing cataract surgery. The charts were reviewed to investigate the failure rate of axial length (AL) measurement of the SS-OCT biometer, partial coherence interferometry (PCI), and A-scan ultrasonography (US) according to cataract type and severity. AL and keratometry in 164 eyes with the same IOL inserted were measured using the SS-OCT biometer, PCI, and A-scan US. The SRK/T formula was used to calculate IOL power. The mean absolute error (MAE) and percentage of eyes with a prediction error (PE) of ±0.50 D were compared. RESULTS: The AL measurement failure rate was 0.00% for A-scan US, 2.32% for the SS-OCT biometer, and 15.31% for PCI. The number of eyes measured using three devices (SS-OCT biometer, PCI, and A-scan US) was 128 (Group A) and the number of eyes measured using two devices (SS-OCT biometer and A-scan US) was 36 (Group B). The score of posterior subcapsular opacity was significantly different between two groups (p < .001). The SS-OCT biometer and PCI showed significantly lower MAE compared to A-scan US in Group A (p = 0.027). Using SS-OCT biometer, MAE showed no significant difference between Group A (0.36 ± 0.27) and Group B (0.36 ± 0.31) (p = 0.785). Whereas, MAE of A-scan US was significantly higher than Group A (0.47 ± 0.39) in Group B (0.64 ± 0.36) (p = 0.023). CONCLUSIONS: Using biometry with advanced OCT is useful in clinical practice as it is more effective in obtaining biometric measurements in the eyes with PSC and provides accurate measurements for IOL power calculation regardless of cataract type and severity. TRIAL REGISTRATION: Retrospectively registered. Registration number: KC16RISI1020 . Registered 03 January 2018.

Inactivation of USP14 Perturbs Ubiquitin Homeostasis and Delays the Cell Cycle in Mouse Embryonic Fibroblasts and in Fruit Fly Drosophila.

BACKGROUND/AIMS: The 26S proteasome is the key proteolytic complex for recognition and degradation of polyubiquitinated target substrates in eukaryotes. Among numerous proteasome-associated proteins, a deubiquitinating enzyme (DUB) USP14 has been identified as an endogenous inhibitor of the proteasome. Here, we explored the complex regulatory functions of USP14 that involve ubiquitin (Ub) homeostasis and substrate degradation in flies and mammals. METHODS: USP14-null primary and immortalized mouse embryonic fibroblasts (MEFs) and USP14 knocked-down Drosophila were analyzed in this study. We measured proteasome and DUB activities using fluorogenic reporter substrates and adduct-forming probes. To examine the levels of ubiquitin, we performed immunoblotting and immunohistochemistry. Mass spectrometry (MS) was used to examine polyUb chain linkages and USP14-interacing proteins. Cell cycle was analyzed by flow cytometry, BrdU labeling, and phospho-histone H3 staining. RESULTS: The homeostasis of Ub in USP14-/-MEFs was markedly perturbed because of facilitated clearance of Ub. This phenomenon was recapitulated in muscles of USP14-deficient Drosophila with old ages. Absolute quantitation using MS also revealed that USP14-/- MEFs contained significantly increased amounts of Ub, compared with wild-type. The key phenotype of USP14-/- MEFs was their delayed proliferation originated from prolonged interphase possibly through aberrant degradation of cyclins A and B1. We found that knocking down USP14 in Drosophila resulted in delayed eye development associated with reduced mitotic activity. CONCLUSION: Our study identifies novel cellular functions of USP14 not only in cellular Ub hometostasis but also in cell cycle progression. USP14 was also essential for proper Drosophila eye development. These results strongly suggest that the USP14-mediated proteasome activity regulation may be directly related to various human diseases including cancer.

Cancer risk in patients with Behçet disease: A nationwide population-based dynamic cohort study from Korea.

BACKGROUND: Many studies have shown a link between inflammation and cancer development. However, there are few studies regarding the correlation between Behçet disease (BD) and cancer. OBJECTIVES: To determine the overall cancer risk and risk for specific cancers in patients with BD. METHODS: Patients with BD (n = 14,137; mean age, 44.2 ± 12.5 years; male patients, 32.4%) without known previous cancer were selected from the Korean National Health Insurance Database between 2007 and 2014. An age- and sex-matched control population of individuals without BD was randomly sampled at a ratio of 10:1. Both cohorts were followed for incident cancer until 2015. RESULTS: Overall, cancer was newly diagnosed in 451 patients with BD (3.19%) and 3975 controls (2.81%) during the follow-up period. Patients with BD showed a significantly higher risk for cancer compared with the controls (hazard ratio [HR], 1.134; 95% confidence interval [CI], 1.029-1.25), leukemia (HR, 5.801; 95% CI, 3.24-10.385), lymphoma (HR, 2.584; 95% CI, 1.559-4.283), oral cavity and pharyngeal cancer (HR, 2.113; 95% CI, 1.102-4.052), thyroid cancer (HR, 1.256; 95% CI, 1.05-1.501), and prostate cancer (HR, 1.784; 95% CI, 1.141-2.791). LIMITATIONS: The treatment or severity of diseases in each individual was not examined. CONCLUSIONS: Patients with BD had a higher risk for overall cancer compared with controls without BD. Physicians should carefully monitor patients with BD for the potential development of malignancies.

Incidence and Prognosis of Subsequent Cholangiocarcinoma in Patients with Hepatic Resection for Bile Duct Stones.

BACKGROUND AND AIM: Cholangiocarcinoma (CCA) often develops after the hepatic resection for hepatolithiasis as well as indwelling it. We studied the incidence and prognosis of subsequent CCA in patients with hepatolithiasis in South Korea. METHODS: We identified individuals with diagnosed CCA at the time of or after surgery, during 2002-2016, from the Korean National Health Insurance. The incidences and survival rates of subsequent CCA were analyzed and compared with concomitant CCA. The standardized incidence ratios (SIRs) of CCA in this cohort were evaluated in the standard Korean population. All data were stratified by the presence of intrahepatic or extrahepatic CCA, age and sex. RESULTS: Of the 7852 patients with hepatectomy for BDS, 433 (5.84%) had concomitant CCA. Over the 12-year follow-up, 107 of 7419 (1.98%) patients were diagnosed with subsequent CCA. Patients with hepatic resection for BDS revealed higher SIRs for subsequent CCA (12.89, 95% CI 10.96-15.15) in cases of both intrahepatic CCA (13.40, 10.55-17.02) and extrahepatic CCA (12.42, 9.98-15.46). The median survival time for subsequent CCA was 0.87 years, while that for concomitant CCA was 2.79 years. Having subsequent CCA (HR 2.71, 95% CI 2.17-3.40) and being male (HR 1.28, 1.05-1.57) were related to a shorter survival time. The CCA site and age at CCA diagnosis were not related to prognoses. CONCLUSIONS: Subsequent CCA developed in 2% of the patients with hepatic resection for benign BDS until 10 years and was associated with poorer prognoses than concomitant CCA. Future studies focused on the long-term surveillance for CCA in such patients are needed.

Disease-Associated Mutations of TREM2 Alter the Processing of N-Linked Oligosaccharides in the Golgi Apparatus.

The triggering receptor expressed on myeloid cells 2 (TREM2) is an immune-modulatory receptor involved in phagocytosis and inflammation. Mutations of Q33X, Y38C and T66M cause Nasu-Hakola disease (NHD) which is characterized by early onset of dementia and bone cysts. A recent, genome-wide association study also revealed that single nucleotide polymorphism of TREM2, such as R47H, increased the risk of Alzheimer's disease (AD) similar to ApoE4. However, how these mutations affect the trafficking of TREM2, which may affect the normal functions of TREM2, was not known. In this study, we show that TREM2 with NHD mutations are impaired in the glycosylation with complex oligosaccharides in the Golgi apparatus, in the trafficking to plasma membrane and further processing by γ-secretase. Although R47H mutation in AD affected the glycosylation and normal trafficking of TREM2 less, the detailed pattern of glycosylated TREM2 differs from that of the wild type, thus suggesting that precise regulation of TREM2 glycosylation is impaired when arginine at 47 is mutated to histidine. Our results suggest that the impaired glycosylation and trafficking of TREM2 from endoplasmic reticulum/Golgi to plasma membrane by mutations may inhibit its normal functions in the plasma membrane, which may contribute to the disease.

Nitric oxide induction of Parkin translocation in PTEN-induced putative kinase 1 (PINK1) deficiency: functional role of neuronal nitric oxide synthase during mitophagy.

The failure to trigger mitophagy is implicated in the pathogenesis of familial Parkinson disease that is caused by PINK1 or Parkin mutations. According to the prevailing PINK1-Parkin signaling model, mitophagy is promoted by the mitochondrial translocation of Parkin, an essential PINK1-dependent step that occurs via a previously unknown mechanism. Here we determined that critical concentrations of NO was sufficient to induce the mitochondrial translocation of Parkin even in PINK1 deficiency, with apparent increased interaction of full-length PINK1 accumulated during mitophagy, with neuronal nitric oxide synthase (nNOS). Specifically, optimum levels of NO enabled PINK1-null dopaminergic neuronal cells to regain the mitochondrial translocation of Parkin, which appeared to be significantly suppressed by nNOS-null mutation. Moreover, nNOS-null mutation resulted in the same mitochondrial electron transport chain (ETC) enzyme deficits as PINK1-null mutation. The involvement of mitochondrial nNOS activation in mitophagy was further confirmed by the greatly increased interactions of full-length PINK1 with nNOS, accompanied by mitochondrial accumulation of phospho-nNOS (Ser(1412)) during mitophagy. Of great interest is that the L347P PINK1 mutant failed to bind to nNOS. The loss of nNOS phosphorylation and Parkin accumulation on PINK1-deficient mitochondria could be reversed in a PINK1-dependent manner. Finally, non-toxic levels of NO treatment aided in the recovery of PINK1-null dopaminergic neuronal cells from mitochondrial ETC enzyme deficits. In summary, we demonstrated the full-length PINK1-dependent recruitment of nNOS, its activation in the induction of Parkin translocation, and the feasibility of NO-based pharmacotherapy for defective mitophagy and ETC enzyme deficits in Parkinson disease.

Single octapeptide deletion selectively processes a pathogenic prion protein mutant on the cell surface.

The number of octapeptide repeats has been considered to correlate with clinical and pathogenic phenotypes of prion diseases resulting from aberrant metabolism of prion protein (PrP). However, it is still poorly understood how this motif affects PrP metabolism. Here, we discover homozygous single octapeptide repeat deletion mutation in the PRNP gene encoding PrP in HeLa cells. The level of PrP proves to be unaffected by this mutation alone, but selectively reduced by additional pathogenic mutations within internal hydrophobic region of PrP. The pattern and relative amount of newly synthesized A117V mutant is unaffected, whereas the mutant appears to be differentially distributed and processed on the cell surface by single octapeptide deletion. This study provides an insight into a novel mutant-specific metabolism of PrP on the cell surface.

N-terminal pro-B-type natriuretic peptide and cardiac troponin T in non-immune hydrops.

AIM: We analyzed the N-terminal pro-B-type natriuretic peptide (NT-proBNP) and cardiac troponin T (cTnT) in non-immune hydrops cases, to determine whether congestive heart failure or inadequate cardiac output may be mechanisms underlying abnormal fluid collection in hydrops. METHODS: The study population consisted of singleton neonates with non-immune hydrops and healthy control neonates, matched for both gestational age at delivery and mode of delivery. The subjects were divided into three groups: group 1, hydrops of cardiac origin (n=6); group 2, hydrops of non-cardiac origin (n=17); and group 3, control (n=23). The NT-proBNP and cTnT were measured in the cord blood collected at the time of delivery. RESULTS: The median cord blood concentrations of NT-proBNP and cTnT were not different between the hydrops (groups 1 and 2) or control cases. However, the NT-proBNP level was elevated in group 1 compared with groups 2 and 3. The concentrations of cTnT did not differ among the three groups. CONCLUSIONS: The concentration of NT-proBNP was only elevated in cases of hydrops of cardiac origin. Congestive heart failure might not be the underlying mechanism of fluid collection in cases of non-immune hydrops of non-cardiac origin, and NT-proBNP may be a promising tool to differentiate the origin of hydrops.