Crosstalk between mucosal microbiota, host gene expression, and sociomedical factors in the progression of colorectal cancer.

Various omics-based biomarkers related to the occurrence, progression, and prognosis of colorectal cancer (CRC) have been identified. In this study, we attempted to identify gut microbiome-based biomarkers and detect their association with host gene expression in the initiation and progression of CRC by integrating analysis of the gut mucosal metagenome, RNA sequencing, and sociomedical factors. We performed metagenome and RNA sequencing on colonic mucosa samples from 13 patients with advanced CRC (ACRC), 10 patients with high-risk adenoma (HRA), and 7 normal control (NC) individuals. All participants completed a questionnaire on sociomedical factors. The interaction and correlation between changes in the microbiome and gene expression were assessed using bioinformatic analysis. When comparing HRA and NC samples, which can be considered to represent the process of tumor initiation, 28 genes and five microbiome species were analyzed with correlation plots. When comparing ACRC and HRA samples, which can be considered to represent the progression of CRC, seven bacterial species and 21 genes were analyzed. When comparing ACRC and NC samples, 16 genes and five bacterial species were analyzed, and four correlation plots were generated. A network visualizing the relationship between bacterial and host gene expression in the initiation and progression of CRC indicated that Clostridium spiroforme and Tyzzerella nexilis were hub bacteria in the development and progression of CRC. Our study revealed the interactions of and correlation between the colonic mucosal microbiome and host gene expression to identify potential roles of the microbiome in the initiation and progression of CRC. Our results provide gut microbiome-based biomarkers that may be potential diagnostic markers and therapeutic targets in patients with CRC.

Detection of Microsatellite Instability in Colorectal Cancer Patients With a Plasma-Based Real-Time PCR Analysis.

A microsatellite instability (MSI) test is crucial for screening for HNPCC (Hereditary nonpolyposis colorectal cancer; Lynch syndrome) and optimization of colorectal cancer (CRC) treatment. Mismatch repair (MMR) deficiency is a predictor for good response of immune checkpoint inhibitors in various malignancies. In this study, we evaluated the results of a newly developed plasma-based real-time PCR kit for the detection of MSI in CRC patients. We assessed a peptide nucleotide acid (PNA) probe-mediated real-time PCR test (U-TOP MSI Detection Kit Plus) that determines MSI status by using amplicon melting analysis of five markers (NR21, NR24, NR27, BAT25, and BAT26) from plasma. Eighty-four CRC patients (46 dMMR and 38 pMMR) with colorectal cancer were analyzed. The concordance rate of MSI status assessment between the plasma kit and IHC was 63.0% in dMMR patients (29/46), but in the pMMR evaluation, a 100% (38/38) concordance rate was observed. In the evaluation of the performance of a custom tissue U-TOP MSI Detection Kit and plasma kit in 28 patients, sensitivity, specificity, PPV (positive predictive value) and NPV (negative predictive value) of plasma kit were 68.4, 100, 100, and 44.4%, respectively, with the tissue U-TOP MSI Detection Kit. Our results demonstrate the feasibility of a non-invasive and rapid plasma-based real-time PCR kit (U-TOP MSI Detection Kit Plus) for the detection of MSI in colorectal cancer.

RNA-sequencing identification and validation of genes differentially expressed in high-risk adenoma, advanced colorectal cancer, and normal controls.

Distinct gene expression patterns that occur during the adenoma-carcinoma sequence need to be determined to analyze the underlying mechanism in each step of colorectal cancer progression. Elucidation of biomarkers for colorectal polyps that harbor malignancy potential is important for prevention of colorectal cancer. Here, we use RNA sequencing to determine gene expression profile in patients with high-risk adenoma treated with endoscopic submucosal dissection by comparing with gene expression in patients with advanced colorectal cancer and normal controls. We collected 70 samples, which consisted of 27 colorectal polyps, 24 cancer tissues, and 19 normal colorectal mucosa. RNA sequencing was performed on an Illumina platform to select differentially expressed genes (DEGs) between colorectal polyps and cancer, polyps and controls, and cancer and normal controls. The Kyoto Gene and Genome Encyclopedia (KEGG) and gene ontology (GO) analysis, gene-concept network, GSEA, and a decision tree were used to evaluate the DEGs. We selected the most highly expressed genes in high-risk polyps and validated their expression using real-time PCR and immunohistochemistry. Compared to patients with colorectal cancer, 82 upregulated and 24 downregulated genes were detected in high-risk adenoma. In comparison with normal controls, 33 upregulated and 79 downregulated genes were found in high-risk adenoma. In total, six genes were retrieved as the highest and second highest expressed in advanced polyps and cancers among the three groups. Among the six genes, ANAX3 and CD44 expression in real-time PCR for validation was in good accordance with RNA sequencing. We identified differential expression of mRNAs among high-risk adenoma, advanced colorectal cancer, and normal controls, including that of CD44 and ANXA3, suggesting that this cluster of genes as a marker of high-risk colorectal adenoma.

Ethanol-dispersed and antibody-conjugated polymer nanofibers for the selective capture and 3-dimensional culture of EpCAM-positive cells.

Electrospun and ethanol-dispersed polystyrene-poly(styrene-co-maleic anhydride) (PS-PSMA) nanofibers (NFs) were used as a platform for the selective capture and three-dimensional culture of EpCAM-positive cells in cell culture medium and whole blood. The NFs were treated with streptavidin to facilitate bond formation between the amino groups of streptavidin and the maleic anhydride groups of the NFs. A biotinylated anti-EpCAM monoclonal antibody (mAb) was attached to the streptavidin-conjugated NFs via the selective binding of streptavidin and biotin. Upon simple mixing and shaking with EpCAM-positive cancer cells in a wide concentration range from 10 to 1000,000 cells per 10mL, the mAb-attached NFs (mAb-NFs) captured the Ep-CAM positive cells in an efficiency of 59%-67% depending on initial cell concentrations, with minor mechanical capture of 14%-36%. Captured cells were directly cultured, forming cell aggregates, in the NF matrix, which ensures the cell proliferation and follow-up analysis. Furthermore, the capture capacity of mAb-NFs was assessed in the presence of whole blood and blood lysates, indicating cluster formation that captured target cells. It is anticipated that the antibody-attached NFs can be employed for the capture and analysis of very rare EpCAM positive circulating cancer cells.

Experiences of violence, burnout and job satisfaction in Korean nurses in the emergency medical centre setting.

The aim of this study was to examine the experience of violence in relation to burnout and job satisfaction in Korean nurses in the emergency medical centre setting. Participants were 236 nurses in the emergency medical centre setting of three metropolitan areas in Korea. Measures included a general characteristics form, characteristics related to experiences of violence, the Maslach Burnout Inventory and the Minnesota Satisfaction Questionnaire. Data were collected from June 2013 to February 2014. In the prediction model, 33.4% of burnout was explained and 35.7% for job satisfaction. The greatest influence on burnout was handling violence, followed by verbal abuse. The greatest influence on job satisfaction was physical threat, followed by handling violence. The study shows that burnout and job satisfaction of Korean nurses in the emergency medical centre setting are related to experiences of violence such as verbal abuse, physical threat and physical violence, as well as handling violence.

[Adaptation Process to Group Home Living by Older Adults].

PURPOSE: The purpose of the study was to explore and describe the adaptation process of older people to group homes. METHODS: Participants were twenty older adults aged 65 or older who were living in group homes. Data were collected from January to April, 2015. In-depth unstructured interviews were conducted with individual participants. Data were analyzed using Strauss and Corbin's grounded theory method. RESULTS: From open coding, 100 concepts, 38 sub-categories, and 14 categories were identified. Analysis showed that the central phenomenon of the adaptation process of older people to group homes was 'gradually giving up'. Causal conditions were 'good-for-nothing body', contextual conditions were 'pushed', 'beleaguered'. Intervening conditions were 'reliable pillar: children', 'having affection more than having it from family: facility workers', 'comfort - like feeling at home', 'relieved: system'. Action/interaction strategies were 'facing the unfamiliar reality', 'building relationships with other people', 'accepting reality'. Consequences were 'a good place, more than expected', 'hope for the remaining days', 'waiting for a peaceful death'. CONCLUSION: The results of this study provide an in-depth understanding of the experience of the adaptation process of older people to group homes. The findings from this study can be used as basic data to establish policies to increase the number of small scale facilities which can help older adults adapt easily to the facilities.

Alternative purification method for recombinant measles viral nucleoprotein expressed in insect cells by ion-exchange chromatography.

Recombinant measles virus nucleoproteins (rMeV N) and fusion (F) proteins were characterized as major antigenic proteins expressed in insect cells mediated by recombinant baculoviruses (rBVs). Band intensities were analyzed by Western blotting to recognize IgG and IgM antibodies against the rMeV N and F proteins in human sera and cerebrospinal fluids (CSFs) from patients with measles infections. Positive results from the blots using the rMeV N were consistent with the results of enzyme-linked immunosorbent assays (ELISAs) in which whole viral proteins were used as antigens. Human sera and CSFs reacted more strongly with the rMeV N than with the rMeV F proteins prepared in an identical expression system. For efficient and reliable purification, ion-exchange chromatography using Source Q anion resin was applied, and high-purity rMeV N protein was harvested. To characterize the similarity with the native viral protein to purified N protein, structural mimicry of purified recombinant proteins with intact rMeV N was shown through transmission electron microscopy, and the truncation and the phosphorylation status of the expressed protein were analyzed. These results suggest that the rMeV N purified by ion-exchange chromatography has features similar to those of naïve N including a self-assembled structure, phosphorylation and antigenic function. Thus, these expression and purification methods can be applied to the large-scale production of the rMeV N, which is essential for the development of new diagnostic tools and vaccines for acute and chronic MeV infections.

Prevalence and molecular epidemiology of human coronavirus HKU1 in patients with acute respiratory illness.

In 2005, human coronavirus HKU1 (HCoV-HKU1) was isolated and identified from a 71-year-old man with pneumonia in Hong Kong. To identify and classify genotypes of HCoV-HKU1 in Korea, a sensitive, specific, and quantitative real-time polymerase chain reaction (PCR) assay was developed and analyzed the sequences of HCoV-HKU1 isolated in Korea. A total of 1,985 respiratory specimens taken from patients with acute respiratory illness were tested for HCoV-HKU1 from January 2007 to May 2008. The major clinical symptoms associated with HCoV-HKU1 infection were examined statistically and sequence variations of the RNA-dependent RNA polymerase (RdRp), spike, and nucleocapsid genes were also analyzed. Fifty cases (2.5%) HCoV-HKU1 were identified by real-time PCR and viral loads ranged from 6.7 × 10(4) to 1.6 × 10(9) copies/ml. The clinical symptoms of HCoV-HKU1 infection included rhinorrhea (72%), cough (64%), nasal congestion (56%), fever (32%), sputum (30%), sore throat (18%), chills (16%), postnasal discharge (14%), and tonsillar hypertrophy (10%). There was a seasonal distribution of HCoV-HKU1 infection, peaking in winter and spring. Both genotypes A and B were detected but no recombination between them was found. This is the first report on the identification and genotyping of HCoV-HKU1 as a causative agent of acute respiratory illness in Korea. The data suggest that at least two genotypes, A and B, of HCoV-HKU1 with scattered silent mutations were circulating in Korea from 2007 to 2008.

Evaluation of an expanded case definition for vaccine-modified measles in a school outbreak in South Korea in 2010.

In this study, we have described the clinical characteristics of vaccine-modified measles to assess the performance of an expanded case definition in a school outbreak that occurred in 2010. The sensitivity, specificity, and the positive and negative predictive values were evaluated. Among 74 cases of vaccine-modified measles, 47 (64%) met the original case definition. Fever and rash were observed in 73% (54/74); fever was the most common (96%, 71/74) presenting symptom, and rash was noted in 77% (57/74) of the cases. The original case definition showed an overall sensitivity of 63.5% and a specificity of 100.0%. The expanded case definition combining fever and rash showed a higher sensitivity (72.9%) but a lower specificity (88.2%) than the original. The presence of fever and one or more of cough, coryza, or conjunctivitis scored the highest sensitivity among the combinations of signs and symptoms (77.0%), but scored the lowest specificity (52.9%). The expanded case definition was sensitive in identifying suspected cases of vaccine-modified measles. We suggest using this expanded definition for outbreak investigation in a closed community, and consider further discussions on expanding the case definition of measles for routine surveillance in South Korea.

Seroprevalence of measles among children affected by national measles elimination program in Korea, 2010.

BACKGROUND: Following the implementation of national measles elimination plan in Korea, the elimination was declared in 2006. In order to sustain the elimination, high population immunity should be continuously monitored. To evaluate the current age-related susceptibility within the Korean population, we conducted the seroprevalence in children and adolescents who were affected by the national measles elimination plan. METHODS: We used residual serum specimens to measure measles specific IgG and geometric mean titer (GMT) in birth cohorts 2007-2008 and 1997-2003. Among birth cohorts, 2007-2008 cohorts were grouped to evaluate the timeliness of first dose of MMR, 1994-2003 cohorts were grouped to evaluate the effect of keep-up MMR2 campaign, and 1992-1993 cohorts were grouped to evaluate the effect of catch-up campaign in 2001. RESULTS: Overall, measles seropositivity rate was 86%. The highest seroprevalence of measles IgG was in birth cohorts 2007-2008. Measles seropositivity declined continuously in age groups. The birth cohorts 1994-1996 showed significantly lower levels of seropositivity and GMT than did the other birth cohorts. CONCLUSION: Despite efforts to eliminate measles for the past 10 years in Korea, our study revealed specific birth cohorts remaining at risk for transmission. The adolescents born during 1994-1996 had the lowest measles seropositivity levels, and might represent a 'pocket' that has potential at increased risk for measles transmission. Further discussion for follow-up immunization should be placed for consideration in the near future.

Outbreak of measles in the Republic of Korea, 2007: importance of nosocomial transmission.

BACKGROUND: From 2002 through 2006, Republic of Korea conducted extensive measles elimination activities and declared elimination in 2006. An outbreak of measles involving 180 confirmed cases occurred during 2007. METHODS: An outbreak investigation was performed and enhanced surveillance was implemented. Detailed case investigations and laboratory testing included serologic and molecular diagnostic methods. Cases were classified according to World Health Organization and national guidelines. RESULTS: During 2007, 451 suspected cases were reported and 180 (40%) cases were confirmed as measles during epidemiologic weeks 14-42. Incidence during the outbreak was 3.7 cases per million persons, excluding imported cases. Most confirmed cases were reported from Seoul; 137 (76%) cases were among children <24 months old, 124 (69%) case patients had no history of measles vaccination, and 81 (45%) case patients resulted from nosocomial transmission in 6 hospitals. Community members, patients, and health care workers all contributed to measles virus transmission. Limited outbreak control measures were implemented; high population immunity likely accounted for the self-limited transmission during this outbreak. CONCLUSIONS: Limited outbreaks of measles, in which nosocomial transmission can play an important role, may occur after countries have declared elimination. Timely and opportunistic vaccination may help prevent such outbreaks; high-quality surveillance is critical for their detection.