Open-top Bessel beam two-photon light sheet microscopy for three-dimensional pathology.

Nondestructive pathology based on three-dimensional (3D) optical microscopy holds promise as a complement to traditional destructive hematoxylin and eosin (H&E) stained slide-based pathology by providing cellular information in high throughput manner. However, conventional techniques provided superficial information only due to shallow imaging depths. Herein, we developed open-top two-photon light sheet microscopy (OT-TP-LSM) for intraoperative 3D pathology. An extended depth of field two-photon excitation light sheet was generated by scanning a nondiffractive Bessel beam, and selective planar imaging was conducted with cameras at 400 frames/s max during the lateral translation of tissue specimens. Intrinsic second harmonic generation was collected for additional extracellular matrix (ECM) visualization. OT-TP-LSM was tested in various human cancer specimens including skin, pancreas, and prostate. High imaging depths were achieved owing to long excitation wavelengths and long wavelength fluorophores. 3D visualization of both cells and ECM enhanced the ability of cancer detection. Furthermore, an unsupervised deep learning network was employed for the style transfer of OT-TP-LSM images to virtual H&E images. The virtual H&E images exhibited comparable histological characteristics to real ones. OT-TP-LSM may have the potential for histopathological examination in surgical and biopsy applications by rapidly providing 3D information.

Disparity in the dental care of children and adolescents with autism spectrum disorder in Korea: a national population-based cross-sectional study.

Individuals with autism spectrum disorders (ASD) have difficulty accessing dental care. We aim to verify whether individuals with ASD are properly managed by checking the frequency of dental visits, cost and severity of dental treatment compared with those without ASD. This cross-sectional study used the Korean Health Insurance Database to analyze the frequency, cost and severity of dental treatment in 209,780 people under the age of 19 with or without ASD in 2020. The average frequency of dental visits for individuals without ASD was 2.98 times, which was significantly higher (p < 0.001) than the 2.89 times for those with ASD. However, the average dental cost for individuals with ASD was USD 132.63, which was significantly higher (p < 0.001) than USD 116.57 for those without ASD. Additionally, the average number of times that individuals without ASD received severe dental treatment was 1.23 times, significantly higher than the 1.15 times for those with ASD. Further, per 10,000 people, we found that trauma treatment was recorded for an average of 21.90 individuals with ASD, significantly higher than the 7.75 recorded for those without ASD (p < 0.001). Individuals with ASD encounter significant disparities in accessing dental care, as evidenced by their relatively infrequent dental visits. This discrepancy can be attributed to various barriers including the financial burden compared with those without ASD.

Approval status of essential therapeutic drugs for systemic sclerosis versus that of drugs for rheumatoid arthritis.

Objective: Systemic sclerosis, a rare disease characterized by chronic multisystem fibrosis, requires lifelong management, necessitating enough insurance coverage for the patient. Official drug approval is the first step to ensuring that the drug is covered by insurance. In this study, we investigated the approval status of essential therapeutic drugs for systemic sclerosis across eight countries and compared it with that of drugs for rheumatoid arthritis. Methods: The essential therapeutic drug lists for systemic sclerosis and rheumatoid arthritis were taken from the guidelines of the American College of Rheumatology and the European Alliance of Associations for Rheumatology. Official drug approval status for the selected drugs was confirmed by searching representative Internet databases from eight countries: the United States, the United Kingdom, Germany, France, Italy, Switzerland, Japan, and the Republic of Korea. Results: A total of 21 and 16 drugs were selected for systemic sclerosis and rheumatoid arthritis, respectively. The drug approval rates of the 21 drugs for systemic sclerosis varied among countries. Most drugs used to treat pulmonary arterial hypertension, which were developed recently and are expensive, are approved by most countries; however, most older drugs-which are still essential for management of Raynaud's phenomenon, digital ulcers, interstitial lung disease, and skin fibrosis-are not approved by most countries. By contrast, almost all of the 16 drugs used to treat rheumatoid arthritis, whether old or new, are approved by most countries. Conclusion: Approval rates for drugs used to treat systemic sclerosis, a rare disease, are much lower than those for drugs used to treat rheumatoid arthritis. Thus, approval rates of essential therapeutic drugs for systemic sclerosis need to improve, which will benefit patients by increasing the number of drugs covered by insurance.

Association between smoking status and handgrip strength in Korean male adults: based on Korea National Health and Nutrition Examination Survey 2016-2019.

Background: Smoking is a well-known risk factor of frailty. Handgrip strength (HGS) is highly representative of muscular strength and is used in the diagnosis of frailty; however, the relationship between smoking and HGS is not clear. We evaluated the relationship between smoking status and HGS. Objectives: This study aimed to evaluate the association between HGS and smoking status. Methods: We enrolled adult males between the ages of 19 and 80 years who participated in the Korea National Health and Nutrition Examination Survey VII-VIII. A chi-square test and ANOVA were performed to compare the mean handgrip strength (mean HGS) between non-smokers, ex-smokers, and current smokers. Logistic regression analysis was performed to determine the association between the smoking status and mean HGS, and additional analyses were performed by dividing subgroups by age. Results: A total of 7,649 participants were analyzed. When the mean HGS and mean dominant HGS were compared according to smoking status, HGS was higher in the right hand (value of p = 0.03) and left hand (value of p < 0.001) in the order of current smokers, ex-smokers, and non-smokers. Comparing HGS of stronger hands, the mean HGS ex-smokers [aOR, (95% confidence interval): 0.61 (0.46-0.82)] and current smokers: 0.55 (0.38-0.78) was higher than that of non-smokers. When subgroup analysis was performed according to age, current smokers aged >60 years had a higher grip strength than non-smokers. Conclusion: Current smokers had a stronger mean HGS than that of ex-smokers and non-smokers. Current smokers older than 60 years appeared to have a stronger mean HGS than ex-smokers and non-smokers of the same age group.

MicroRNA 29a therapy for CEACAM6-expressing lung adenocarcinoma.

BACKGROUND: Non-coding microRNAs (miRNAs) play critical roles in tumor progression and hold great promise as therapeutic agents for multiple cancers. MicroRNA 29a (miR-29a) is a tumor suppressor miRNA that inhibits cancer cell growth and tumor progression in non-small cell lung cancer. Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), which plays an important role in lung cancer progression, has been identified as a target of miR-29a. Here, we evaluated the therapeutic efficacy of a peptide vector capable of delivering miR-29a intracellularly using the acidic tumor microenvironment in a lung adenocarcinoma xenograft mouse model. METHODS: A miRNA delivery vector was constructed by tethering the peptide nucleic acid form of miR-29a to a peptide with a low pH-induced transmembrane structure (pHLIP) to enable transport of the miRNAs across the plasma membrane. Tumor suppressive effects of pHLIP-miR29a on lung adenocarcinoma development in vivo were assessed using a BALB/c xenograft model injected with A549 cells. RESULTS: Incubation of A549 cells with pHLIP-miR-29a at an acidic pH downregulated endogenous CEACAM6 expression and reduced cell viability. Intravenous injection of the mice with pHLIP-miR-29a inhibited tumor growth by up to 18.1%. Intraperitoneal injection of cisplatin reduced tumor volume by 29.9%. Combined pHLIP-miR-29a + cisplatin treatment had an additive effect, reducing tumor volume up to 39.7%. CONCLUSIONS: Delivery of miR-29a to lung adenocarcinoma cells using a pHLIP-mediated method has therapeutic potential as a unique cancer treatment approach.

Prevalence and severity of COVID-19 among children and adolescents with autism spectrum disorders in the Republic of Korea.

LAY ABSTRACT: It is more difficult to prevent coronavirus disease 2019 in children and adolescents with autism spectrum disorder, as they have trouble communicating and adjusting to their new daily lives like wearing masks and social distancing. However, there have not been many studies that focused on coronavirus disease 2019 among children and adolescents with autism spectrum disorder. We included all Korean citizens under the age of 19 as our study subjects. Among them, we found out the prevalence, severity, and case fatality of coronavirus disease 2019 in children and adolescents with and without autism spectrum disorder. The prevalence of coronavirus disease 2019 among children and adolescents with autism spectrum disorder was lower than that of those without autism spectrum disorder. For severity, children and adolescents with autism spectrum disorder were more likely to enter severe stages of disease and had higher hospitalization rates than those without autism spectrum disorder. There were no deaths among children and adolescents with autism spectrum disorder, while a few died among children and adolescents without autism spectrum disorder. However, due to the small number of deaths, it was difficult to determine whether there was a link between autism spectrum disorder and coronavirus disease 2019 deaths. We found that the appropriate quarantine policies have played a great role in sustaining overall low prevalence and higher hospitalization rates among children and adolescents with autism spectrum disorder than those without autism spectrum disorder. Furthermore, because Korea has fewer schools and facilities (i.e. personal care, social training, and skilled nursing facility) for children and adolescents with autism spectrum disorder than other countries, those with autism spectrum disorder have fewer social contacts than even before the COVID-19 pandemic.

Gold single-atoms confined at the CeOx-TiO2interfaces with enhanced low-temperature activity toward CO oxidation.

We use CeOx-TiO2hetero-interfaces generated on the surface of CeOx-TiO2hybrid oxide supporting powders to stabilize Au single-atoms (SAs) with excellent low-temperature activity toward CO oxidation. Based on intriguing density functional theory calculation results on the preferential formation of Au-SAs at the CeOx-TiO2interfaces and the high activity of Au-SAs toward the Mars-van Krevelen type CO oxidation, we synthesized a Au/CeOx-TiO2(ACT) catalyst with 0.05 wt.% of Au content. The Au-SAs stabilized at the CeOx-TiO2interfaces by electronic coupling between Au and Ce showed improved low-temperature CO oxidation activity than the conventional Au/TiO2control group catalyst. However, the light-off profile of ACT showed that the early activated Au-SAs are not vigorously participating in CO oxidation. The large portion of the positive effect on the overall catalytic activity from the low activation energy barrier of ACT was retarded by the negative impact from the decreasing active site density at high temperatures. We anticipate that the low-temperature activity and high-temperature stability of Au-SAs that stand against each other can be optimized by controlling the electronic coupling strength between Au-SAs and oxide clusters at the Au-oxide-TiO2interfaces. Our results show that atomic-precision interface modulation could fine-tune the catalytic activity and stability of Au-SAs.

The impact of the COVID-19 pandemic in the healthcare utilization in Korea: Analysis of a nationwide survey.

BACKGROUND: COVID-19 has brought changes in daily life and increased the medical burden. This study aims to evaluate the delays in healthcare services and related factors in the general population during the COVID-19 pandemic. METHODS: We took a nationally representative sample and conducted a mobile phone-based survey. The study was conducted anonymously. Of the 3377 subjects who consented to participate, a total of 2097 finished the survey. The primary outcome was respondents' experiences with delayed (1) health screenings, (2) non-urgent medical visits, (3) medical visits for chronic disease, and (4) emergency visits during the COVID-19 pandemic. RESULTS: Of 2097 respondents, females, residents of the Seoul metropolitan area, those with private insurance, those without chronic diseases, smokers, and drinkers had higher risk of delays in health screening and non-urgent medical visits after adjustment. Among chronic disease patients, those who were over 60 years old (adjusted odds ratio 0.36, 95% CI 0.14-0.92) showed lower risk of delayed medical visit. Residents of the Seoul metropolitan area, those with private insurance, smokers, and drinkers were all associated with experiencing delayed health screening and non-urgent medical visits had higher risk of delays in chronic disease visits and emergent medical visits. CONCLUSIONS: Delayed access to healthcare services is associated with poor outcomes and may cause different complications. Efforts are needed to prevent delays in medical use due to infectious diseases such as COVID-19. Considering the possibility of the emergence of infectious diseases, various countermeasures are needed to prevent delays in medical visit.

Identifying patients with deteriorating generalized pustular psoriasis: Development of a prediction model.

Generalized pustular psoriasis (GPP) is a life-threatening condition; however, little is known about the factors that can predict GPP patients manifesting a deteriorating course. To investigate the demographics and clinical features of adult inpatient GPP and propose a prediction model for detecting fatal GPP (fGPP) and GPP requiring intensive care unit admission (iGPP) patients, a nationwide population-based retrospective cohort study was conducted. The adult inpatients with GPP from January 2007 to December 2020 were assessed. The 800 cases were aged 51.0 years (median [interquartile range, 37.0-64.0]). Overall, 21 iGPP (64.0 years [54.0-77.0]) and 17 fGPP (75.0 years [68.0-77.0]) cases were identified as deteriorating GPP. Renal disease (odds ratio [OR], 7.31), myocardial infarction history (OR, 4.29), liver disease (OR, 2.82), and diabetes mellitus (OR, 2.34) were identified as predictors for iGPP. For fGPP, myocardial infarction history (OR, 5.10) and psoriasis history (OR, 3.13) were established as predictors. A prediction model with scores ranging 0-11 points showed a reliable diagnostic value in detecting deteriorating GPP (area under the curve = 0.75 for iGPP and 0.83 for fGPP). In conclusion, this study provides the clinical features of deteriorating GPP. A prediction model may help physicians to identify patients with deteriorating GPP.

Interspersing CeOx Clusters to the Pt-TiO2 Interfaces for Catalytic Promotion of TiO2-Supported Pt Nanoparticles.

We propose an interface-engineered oxide-supported Pt nanoparticle-based catalyst with improved low-temperature activity toward CO oxidation. By wet-impregnating 1 wt % Ce on TiO2, we synthesized hybrid oxide support of CeOx-TiO2, in which dense CeOx clusters formed on the surface of TiO2. Then, the Pt/CeOx-TiO2 catalyst was synthesized by impregnating 2 wt % Pt on the CeOx-TiO2 supporting oxide. Pt-CeOx-TiO2 triphase interfaces were eventually formed upon impregnation of Pt on CeOx-TiO2. The Pt-CeOx-TiO2 interfaces open up the interface-mediated Mars-van Krevelen CO oxidation pathway, thus providing additional interfacial reaction sites for CO oxidation. Consequently, the specific reaction rate of Pt/CeOx-TiO2 for CO oxidation was increased by 3.2 times compared with that of Pt/TiO2 at 140 °C. Our results demonstrate a widely applicable and straightforward method of catalytic activation of the interfaces between metal nanoparticles and supporting oxides, which enabled fine-tuning of the catalytic performance of oxide-supported metal nanoparticle classes of heterogeneous catalysts.

The general public's perspectives on telemedicine during the COVID-19 pandemic in Korea: analysis of a nationwide survey.

OBJECTIVES: We investigated the awareness, experience, approval, intention to use, and the desired type of telemedicine among Korean general public. METHODS: From November to December 2020, we conducted an online self-reported survey on awareness, experience, approval, and intent to use telemedicine services among Korean residents aged 20 years or older. A total of 2,097 participants completed the survey. RESULTS: Of the 2,097 participants, 1,558 (74.3%) were aware of, 1,198 (57.1%) approved of, and 1,474 (70.3%) had the intention to use telemedicine. Participants from regions other than the Seoul metropolitan area and Daegu-Gyeongbuk Province (adjusted odds ratio [aOR], 1.29; 95% confidence interval [CI], 1.02 to 1.63), households with a monthly household income of US$6,000 or more (aOR, 1.44; 95% CI, 1.01 to 2.08), participants who had a college/university or associate's degree (aOR, 1.35. 95% CI, 1.04 to 1.75) or a master's degree or above (aOR, 1.73; 95% CI, 1.20 to 2.50), and housewives (aOR, 1.30; 95% CI, 1.03 to 1.64) had higher odds of approval. Elderly participants, those with a chronic disease (aOR, 1.26; 95% CI, 1.04 to 1.54), those who had experienced delays of healthcare services (aOR, 1.94; 95% CI, 1.27 to 2.96), and those who had experience with telemedicine (aOR, 4.28; 95% CI, 1.69 to 10.82) were more likely to intend to use telemedicine services. Regarding types of telemedicine, teleconsultation between doctors showed the highest approval rate (73.1%). CONCLUSIONS: In the context of the coronavirus disease 2019 pandemic, more than 70% of participants had already used or intended to use telemedicine at some point. Groups with a substantial need for telemedicine were more in favor of telemedicine.

Prevalence and Premature Mortality Statistics of Autism Spectrum Disorder Among Children in Korea: A Nationwide Population-Based Birth Cohort Study.

BACKGROUND: The aim of this study was to estimate the 8-year prevalence and mortality statistics of autism spectrum disorder (ASD) according to birth year (2002-2012). METHODS: We used the National Health Insurance Service database with 4,989,351 children born from 2002 to 2012 including 35,529 children diagnosed with ASD until 8 years of age. The 8-year cumulative prevalence of ASD was calculated annually (2010-2020) with 8 years of follow-up. The 8-year mortality was estimated using Cox models adjusted for sex, household income, area of residence, and year of birth. RESULTS: Of the 473,494 children born in 2002, 2,467 (5.2 per 1,000 births) were diagnosed with ASD until 2010. The ASD prevalence was 2.6 times higher among boys (1,839; 7.4 per 1,000 boy births) than girls (628; 2.8 per 1,000 girl births). Of the 467,360 children born in 2012, 4,378 (9.4 per 1,000 births) were diagnosed with ASD until 2020. The ASD prevalence was 2.7 times higher among boys (3,246; 13.5 per 1,000 boy births) than girls (1,132; 5.0 per 1,000 girl births). The risk of all-cause mortality was higher among children with ASD than those without (hazard ratio [HR], 2.340; 95% confidence interval [CI], 2.063-2.654), which is substantially higher among girls (HR, 4.223; 95% CI, 3.472-5.135) than boys (HR, 1.774; 95% CI, 1.505-2.090). CONCLUSION: The present study demonstrated that national-level prevalence and mortality statistics of ASD can be estimated effectively using claims data comprising newborns born each year and followed up for to the age of interest. Because this information is essential to establish evidence-based policies, health authorities need to consider producing epidemiological information of ASD continuously using the same methodology.

CD48-expressing non-small-cell lung cancer cells are susceptible to natural killer cell-mediated cytotoxicity.

The susceptibility of cancer cells to natural killer (NK) cell-mediated cytotoxicity depends on the balance of activating and inhibitory ligands expressed on their surface. Although many types of cancer cells are killed by NK cells, non-small-cell lung cancer (NSCLC) cells are relatively resistant to NK cell-mediated cytotoxicity. In this study, we showed that several NSCLC cell lines have differential sensitivity to NK cell-mediated cytotoxicity: NCI-H522 cells were highly sensitive, but A549, NCI-H23, NCI-H1915, and NCI-H1299 were resistant. Among activating ligands such as CD48, HLA-A/B/G, ICAM-1, MICA/B, and ULBPs, only CD48 rendered NCI-H522 cells susceptible to NK cell-mediated cytotoxicity, which was proved by using CD48 siRNA and neutralizing antibody. CD48-positive NCI-H522 cells established a more stable contact with NK cells than did CD48-negative A549 and CD48 siRNA cell-transfected NCI-H522 cells. Taken together, these data demonstrate that CD48-positive NSCLC cells might be susceptible to NK cell-mediated cytotoxicity, which provide information on how to stratify NSCLC patients potentially responsive to NK-cell therapy.

Impact of the COVID-19 Pandemic on the Health Status and Behaviors of Adults in Korea: National Cross-sectional Web-Based Self-report Survey.

BACKGROUND: The COVID-19 pandemic has radically shifted living practices, thereby influencing changes in the health status and behaviors of every person. OBJECTIVE: The aim of this study was to investigate the impact of COVID-19 on the self-reported health status and health behaviors along with any associated factors in adults in the Republic of Korea wherein no stringent lockdown measures were implemented during the pandemic. METHODS: We conducted a web-based self-reported survey from November 2020 to December 2020. The study participants (N=2097) were identified through quota sampling by age, sex, and geographical regions among residents aged 19 years or older in Korea. The survey collected information on basic demographics, changes in self-reported health status, and health behaviors during the COVID-19 pandemic. Self-reported health status and health behaviors were categorized into 3 groups: unchanged, improved, or worsened. A chi-square test and logistic regression analyses were conducted. RESULTS: With regard to changes in the self-reported health status, the majority (1478/2097, 70.5%) of the participants reported that their health was unchanged, while 20% (420/2097) of the participants reported having worser health after the COVID-19 outbreak. With regard to changes in health behaviors, the proportion of participants who increased tobacco consumption was similar to that of those who decreased tobacco consumption (110/545, 20.2% vs 106/545, 19.5%, respectively), while the proportion of those who decreased their drinking frequency was more than twice as many as those who increased their drinking frequency (578/1603, 36.1% vs 270/1603, 16.8%, respectively). Further, those who decreased their exercising frequency were more than those who increased their exercising frequency (333/823, 15.9% vs 211/823, 10%, respectively). The factor that had the greatest influence on lifestyle was age. In the subgroup analysis, the group aged 20-29 years had the highest number of individuals with both a worsened (100/377, 26.5%) and an improved (218/377, 15.7%) health status. Further, individuals aged 20-29 years had greater odds of increased smoking (6.44, 95% CI 2.15-19.32), increased alcohol use (4.64, 95% CI 2.60-8.28), and decreased moderate or higher intensity aerobic exercise (3.39, 95% CI 1.82-6.33) compared to individuals aged 60 years and older. Younger adults showed deteriorated health behaviors, while older adults showed improved health behaviors. CONCLUSIONS: The health status and the behavior of the majority of the Koreans were not found to be heavily affected by the COVID-19 outbreak. However, in some cases, changes in health status or health behavior were identified. This study highlighted that some groups were overwhelmingly affected by COVID-19 compared to others. Certain groups reported experiencing both worsening and improving health, while other groups reported unchanged health status. Age was the most influential factor for behavior change; in particular, the younger generation's negative health behaviors need more attention in terms of public health. As COVID-19 prolongs, public health interventions for vulnerable groups may be needed.

Mortality of Children with Autism Spectrum Disorder Using Data from a Large-Scale Korean National Cohort.

PURPOSE: This study aimed to investigate the association between autism spectrum disorder (ASD) diagnosis and mortality among children using the data from a large-scale national cohort of Korean infants and children. We also explored the association between hearing impairment detected during early infancy and mortality. MATERIALS AND METHODS: We performed a retrospective cohort study using the nationwide claims data of the Republic of Korea. Children born between 2007 and 2014 (n=3598904) were followed up until 2020. Cox proportional hazard models were used to examine the association between ASD diagnosis and mortality among children. Then, in order to evaluate the association between hearing impairment and mortality, Cox proportional hazard models were built using the responses related to hearing impairment asked during the first health screening (at age 4-6 months). RESULTS: Of the 3598904 children born between 2007 and 2014, 32878 children (0.9%) were diagnosed with ASD until the end of 2020. We identified that ASD diagnosis was associated with higher mortality among children [hazard ratio (HR)=2.5, 95% confidence interval (CI): 2.2-2.9]. This association was stronger among girls (HR=4.8, 95% CI: 3.9-5.8) compared to boys (HR=1.9, 95% CI: 1.6-2.2). We found that hearing impairment detected during infancy was associated with higher mortality among children with ASD diagnosis. CONCLUSION: ASD diagnosis was associated with higher mortality among Korean children. This association was stronger in girls than in boys. Hearing impairment detected during infancy was also associated with higher mortality among children diagnosed with ASD.

In Situ Formation of Ag Nanoparticles for Fiber Strain Sensors: Toward Textile-Based Wearable Applications.

Wearable electronic devices have attracted significant attention as important components in several applications. Among various wearable electronic devices, interest in textile electronic devices is increasing because of their high deformability and portability in daily life. To develop textile electronic devices, fiber-based electronic devices should be fundamentally studied. Here, we report a stretchable and sensitive fiber strain sensor fabricated using only harmless materials during an in situ formation process. Despite using a mild and harmless reducing agent instead of typical strong and hazardous reducing agents, the developed fiber strain sensors feature a low initial electrical resistance of 0.9 Ω/cm, a wide strain sensing range (220%), high sensitivity (∼5.8 × 104), negligible hysteresis, and high stability against repeated stretching-releasing deformation (5000 cycles). By applying the fiber sensors to various textiles, we demonstrate that the smart textile system can monitor various gestures in real-time and help users maintain accurate posture during exercise. These results will provide meaningful insights into the development of next-generation wearable applications.

Indole-3-carbinol induces apoptosis-like death in Escherichia coli on different contribution of respective reactive oxygen species.

AIMS: Indole-3-carbinol (I3C) is a natural compound derived from brassica vegetables, displaying antibacterial activity. The study aims to elucidate the antibacterial mode of action(s) induced by indole-3-carbionol in Escherichia coli and enhance the understandings on the respective contribution of each reactive oxygen species (ROS), superoxide anion (O2-), hydrogen peroxide (H2O2), hydroxyl radical (OH-) during the process. MAIN METHODS: The antibacterial activity of I3C was assessed through kinetic assay. The generation of ROS was measured by flow cytometer using H2DCFDA dye, while further analysis of respective contribution was done through application of each scavenger: tiron, thiourea and sodium pyruvate. DNA fragmentation and chromatin condensation were observed by TUNEL and DAPI staining agent. Finally, Annexin V/PI, FITC-VAD-FMK and DiBAC4(3) was applied for detection of apoptosis-like death. KEY FINDINGS: I3C exhibited antibacterial activity in E. coli through accumulation of ROS and DNA damage, eventually leading to apoptosis-like death. Contribution of each ROS displayed respective manner, OH- exerting the most potent influence whereas O2- showed least impact. SIGNIFICANCE: Our study is the first to link I3C to the bacterial apoptosis-like death and displays the potential of this agent as a candidate for potential drugs that could help regulating the E. coli, an opportunistic human pathogen. Moreover, the study focused on investigating the individual contribution of each ROS during the process, trying to enhance the understanding regarding ROS and cellular processes followed by oxidative stress in bacteria.

Bile-derived circulating extracellular miR-30d-5p and miR-92a-3p as potential biomarkers for cholangiocarcinoma.

BACKGROUND: Cholangiocarcinoma (CCA) is from cholangiocytes, and therefore bile is a potentially rich source of biomarkers for CCA. The aim of the study was to identify and validate microRNAs (miRNAs) in bile samples that are differentially expressed between benign biliary disease (BBD) and CCA. METHODS: Bile samples from 106 patients with obstructive biliary disease were allocated consecutively to a discovery set (10 patients with BBD and 11 with CCA) and then a validation set (48 patients with BBD and 37 with CCA). An miRNA microarray platform was used to screen 1209 miRNAs in the discovery set. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) was used to validate the profiling results in the discovery and validation sets. In addition, the levels of carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) were determined from patient serum samples. RESULTS: Microarray profiling showed that miR-30d-5p and miR-92a-3p were significantly upregulated in bile from the CCA group compared with those from the BBD group. qRT-PCR results indicated that the expression levels of miR-30d-5p and of miR-92a-3p were significantly upregulated in the CCA group compared to the BBD group, validating the miRNA microarray results. Pathway analysis suggested that putative target genes of miR-30d-5p and of miR-92a-3p were involved in CCA-associated signalling pathways, such as Hippo, Wnt, p53, MAPK, and EGFR. Receiver operating curve analysis revealed that the areas under the curve for bile miR-30d-5p, miR-92a-3p, serum CA19-9, and CEA were 0.730, 0.652, 0.675, and 0.603, respectively, and bile miR-30d-5p showed the best diagnostic performance with a sensitivity of 81.1% and a specificity of 60.5%. CONCLUSIONS: The levels of extracellular miR-30d-5p and miR-92a-3p in bile were significantly higher in patients with CCA than those in patients with BBD. Bile-derived circulating extracellular miR-30d-5p and miR-92a-3p are potential biomarkers for discriminating CCA from BBD.

Exosomal miR-181b-5p Downregulation in Ascites Serves as a Potential Diagnostic Biomarker for Gastric Cancer-associated Malignant Ascites.

PURPOSE: Peritoneal carcinomatosis in gastric cancer (GC) patients results in extremely poor prognosis. Malignant ascites samples are the most appropriate biological material to use to evaluate biomarkers for peritoneal carcinomatosis. This study identified exosomal MicroRNAs (miRNAs) differently expressed between benign liver cirrhosis-associated ascites (LC-ascites) and malignant gastric cancer-associated ascites (GC-ascites), and validated their role as diagnostic biomarkers for GC-ascites. MATERIALS AND METHODS: Total RNA was extracted from exosomes isolated from 165 ascites samples (73 LC-ascites and 92 GC-ascites). Initially, microarrays were used to screen the expression levels of 2,006 miRNAs in the discovery cohort (n=22). Subsequently, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were performed to validate the expression levels of selected exosomal miRNAs in the training (n=70) and validation (n=73) cohorts. Furthermore, carcinoembryonic antigen (CEA) levels were determined in ascites samples. RESULTS: The miR-574-3p, miR-181b-5p, miR-4481, and miR-181d were significantly downregulated in the GC-ascites samples compared to the LC-ascites samples, and miR-181b-5p showed the best diagnostic performance for GC-ascites (area under the curve [AUC]=0.798 and 0.846 for the training and validation cohorts, respectively). The diagnostic performance of CEA for GC-ascites was improved by the combined analysis of miR-181b-5p and CEA (AUC=0.981 and 0.946 for the training and validation cohorts, respectively). CONCLUSIONS: We identified exosomal miRNAs capable of distinguishing between non-malignant and GC-ascites, showing that the combined use of miR-181b-5p and CEA could improve diagnosis.

Therapeutic Effect of pHLIP-mediated CEACAM6 Gene Silencing in Lung Adenocarcinoma.

Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) plays an important role in lung cancer progression. Here, we examined the therapeutic efficacy of CEACAM6 gene silencing using an siRNA delivery platform targeting the acidic tumour microenvironment in a lung adenocarcinoma xenograft mouse model. An siRNA delivery vector was constructed by tethering the peptide nucleic acid form of an siRNA targeting CEACAM6 (siCEACAM6) to a peptide with a low pH-induced transmembrane structure (pHLIP) to transport siRNAs across the plasma membrane. Specific binding of the pHLIP-siCEACAM6 conjugate to A549 lung adenocarcinoma cells at low pH was demonstrated by flow cytometry. A549 cells incubated with pHLIP-siCEACAM6 at an acidic pH showed downregulated expression of endogenous CEACAM6 protein and reduced cell viability. The in vivo tumour-suppressing effects of pHLIP-siCEACAM6 in lung adenocarcinoma were assessed in a xenograft model generated by injecting BALB/c nude mice with A549 cells. pHLIP-siCEACAM6 treatment alone resulted in tumour growth inhibition of up to 35.5%. When combined with cisplatin treatment, pHLIP-siCEACAM6 markedly enhanced tumour growth inhibition by up to 47%. In conclusion, the delivery of siCEACAM6 to lung adenocarcinoma using the pHLIP peptide has therapeutic potential as a unique cancer treatment approach.