Wastewater Knows Pathogen Spread: Analysis of Residential Wastewater for Infectious Microorganisms including SARS-CoV-2.

BACKGROUND: We aimed to identify the genes of 35 pathogens, including viruses, bacteria, and protozoa that cause waterborne infectious diseases, and to assess the feasibility of a wastewater-based surveillance system. MATERIALS AND METHODS: Wastewater was aseptically sampled once a month from 2 sites. A total of 1 L of wastewater from each site underwent 0.2 µm filtration to generate the sample A. Subsequently, 200 ul of the filtered water was ultra-filtered and concentrated to generate the sample B, which was mixed with sample A in a 1:1 ratio. We performed a Filmarray® Gastrointestinal (GI) panel (BioFire Diagnnostics', Salt Lake City, UT, USA) test to simultaneously detect 13 enterobacteria, 5 enterovirus, and 4 protozoa. RNA was extracted to assess 18 types of viruses. RESULTS: Severe acute respiratory syndrome coronavirus 2 adenovirus, bocavirus, and rhinovirus was detected at both site. Norovirus GI/GII was continuously detected at both sites. Moreover, adenovirus, group A rotaviruses, and hepatitis A virus were frequently detected; however, hepatitis E virus was absent at either site. Campylobacter, enteroaggregative Escherichia coli, enterotoxigenic E. coli, Shiga toxin-producing E. coli, and Clostridioides difficile toxin A/B were detected at both sites. Giardia lamblia was also detected in both sites. CONCLUSION: We analyze multiple infectious disease pathogens under sample surveillance with incidence. Further in-depth studies on wastewater-based surveillance will be feasible and important.

Role of reactive oxygen species in regulating 27-hydroxycholesterol-induced apoptosis of hematopoietic progenitor cells and myeloid cell lines.

Oxysterols are oxygenated derivatives of cholesterol that contain an additional hydroxy, epoxide, or ketone group in the sterol nucleus and/or a hydroxyl group in the side chain of the cholesterol molecule. 27-Hydroxycholesterol (27HC) is a side-chain oxysterol that is oxygenated at the 27th carbon atom of cholesterol. The oxysterol (27HC) is produced via oxidation by sterol 27-hydroxylase (CYP27A1) and metabolized via oxysterol 7a-hydroxylase (CYP7B1) for bile acid synthesis in the liver. A previous study has demonstrated that treatment with the alternative Estrogen receptor alpha (ERα) ligand 27HC induces ERα-dependent hematopoietic stem cell (HSC) mobilization. In addition, Cyp27a1-deficient mice demonstrate significantly reduced 27HC levels and HSC mobilization. Here, we report that exogenous 27HC treatment leads to a substantial reduction in the hematopoietic stem and progenitor cell (HSPC) population owing to significantly increased reactive oxygen species (ROS) levels and apoptosis in the bone marrow (BM). However, 27HC does not influence the population of mature hematopoietic cells in the BM. Furthermore, exogenous 27HC treatment suppresses cell growth and promotes ROS production and apoptosis in leukemic cells. Moreover, acute myeloid leukemia (AML) patients with high CYP7B1 expression (expected to have inhibition of 27HC) had significantly shorter survival than those with low CYP7B1 expression (expected to have an elevation of 27HC). Single-cell RNA-sequencing (scRNA seq) analysis revealed that the expression of CYP7B1 was significantly increased in AML patients. Thus, our study suggests that 27HC may serve as a potent agent for regulating pools of HSPCs and may have an application as a novel therapeutic target for hematological malignancies. Collectively, pharmacological inhibition of CYP7B1 (expected to have an elevation of 27HC) would potentially have fewer long-term hematological side effects, particularly when used in combination with chemotherapy or radiation for the treatment of leukemia patients.

Oxime derivative TFOBO promotes cell death by modulating reactive oxygen species and regulating NADPH oxidase activity in myeloid leukemia.

Several derivatives derived from the oxime structure have been reported as potential anticancer agents in various cancers. Here, we first tested a novel oxime-containing derivative of 2-((2,4,5-trifluorobenzyl)oxy)benzaldehyde oxime (TFOBO) to evaluate its anticancer effect in myeloid leukemic cells. Compared to (2-((2,4,5-trifluorobenzyl)oxy)phenyl)methanol (TFOPM), the oxime derivative TFOBO suppresses leukemic cell growth by significantly increasing reactive oxygen species (ROS) levels and cell death. Leukemic cells treated with TFOBO displayed apoptotic cell death, as indicated by nuclear condensation, DNA fragmentation, and annexin V staining. TFOBO increases Bax/Bcl2 levels, caspase9, and caspase3/7 activity and decreases mitochondrial membrane potential. ROS production was reduced by N-acetyl-L-cysteine, a ROS scavenger, diphenyleneiodonium chloride, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, after exogenous TFOBO treatment. ROS inhibitors protect leukemic cells from TFOBO-induced cell death. Thus, our study findings suggest that TFOBO promotes apoptosis by modulating ROS and regulating NADPH oxidase activity. Collectively, the oxime-containing derivative TFOBO is a novel therapeutic drug for myeloid leukemia.

Microbial and molecular differences according to the location of head and neck cancers.

BACKGROUND: Microbiome has been shown to substantially contribute to some cancers. However, the diagnostic implications of microbiome in head and neck squamous cell carcinoma (HNSCC) remain unknown. METHODS: To identify the molecular difference in the microbiome of oral and non-oral HNSCC, primary data was downloaded from the Kraken-TCGA dataset. The molecular differences in the microbiome of oral and non-oral HNSCC were identified using the linear discriminant analysis effect size method. RESULTS: In the study, the common microbiomes in oral and non-oral cancers were Fusobacterium, Leptotrichia, Selenomonas and Treponema and Clostridium and Pseudoalteromonas, respectively. We found unique microbial signatures that positively correlated with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in oral cancer and positively and negatively correlated KEGG pathways in non-oral cancer. In oral cancer, positively correlated genes were mostly found in prion diseases, Alzheimer disease, Parkinson disease, Salmonella infection, and Pathogenic Escherichia coli infection. In non-oral cancer, positively correlated genes showed Herpes simplex virus 1 infection and Spliceosome and negatively correlated genes showed results from PI3K-Akt signaling pathway, Focal adhesion, Regulation of actin cytoskeleton, ECM-receptor interaction and Dilated cardiomyopathy. CONCLUSIONS: These results could help in understanding the underlying biological mechanisms of the microbiome of oral and non-oral HNSCC. Microbiome-based oncology diagnostic tool warrants further exploration.

Risk stratification of patients with right-sided colorectal cancer based on the tumor-infiltrating M1 macrophage.

The homing of M1 and M2 macrophages may play distinct roles in the tumor microenvironment (TME). However, these roles of macrophages in the TME remain unclear. We downloaded RNA sequencing data from The Cancer Genome Atlas (TCGA) database for patients with CRC. Subsequently, Kaplan-Meier survival curves were generated to assess the differential infiltration of M1 and M2 macrophages based on CRC location. Differentially expressed gene (DEG) and functional analyses were performed to screen the roles of DEGs. Critical prognostic genes were identified using least absolute shrinkage and selection operator regression. The risk scores were calculated for each patient. In patients with right-sided CRC, reduced M1 macrophage infiltration was associated with poor prognosis. M1 macrophage infiltration positively correlated with CD8+ T cell infiltration. A risk model was developed and validated for performance using GSE103479 and GSE72970. Nine genes were identified as independent prognostic genes that could be potential biomarkers for effectively predicting survival in patients with right-sided CRC. Kaplan-Meier curves for overall survival and progression-free survival analyses revealed that the high-risk group of patients with right-sided CRC had a poor prognosis. This novel M1 macrophage-related risk model may provide a gene signature for predicting the survival outcomes of patients with right-sided CRC and facilitate further studies examining the relationship between infiltration of M1 macrophages and the prognosis of such patients.

Investigation of Pathogenic Escherichia coli Contamination of Vegetables Distributed in a Korean Agricultural Wholesale Market.

The purpose of this study was to investigate the monthly contamination rate of pathogenic Escherichia coli, a major cause of food poisoning, in vegetables sold in agricultural wholesale markets, which distribute vegetables from all over the country, in the Incheon Metropolitan City area, South Korea, and to identify a source of the pathogen. In total, 1739 vegetables of 80 types, along with 109 soil, 67 manure, and 33 livestock feces samples, were tested for pathogenic E. coli using polymerase chain reaction, from September 2016 through August 2017. The average annual prevalence rate of vegetables was 5.8%, and the prevalence rate was above 5% from June through October. The highest prevalence rate (15.7%) was recorded in July. Water dropwort showed the highest prevalence rate (28.6%) among the vegetables examined. Pathogenic E. coli was detected in >20 types of the vegetables that were to be consumed without cooking. Among these, the prevalence rates of ponytail radish (n = 21), crown daisy (n = 86), young radish (n = 68), romaine lettuce (n = 133), perilla leaf (n = 103), Korean leek (n = 43), young Chinese cabbage (n = 68), and Chinese cabbage (n = 30) were 9.5%, 8.1%, 7.4%, 6.8%, 4.9%, 4.7%, 4.4%, and 3.3%, respectively. Among the vegetables cooked before consumption, prevalence rates were 28.6%, 27.3%, and 25.0% in wormwood, sweet potato stalk, and edible mountain vegetables (Saussurea sp., etc.), respectively. In soil, manure, and livestock feces, 36.7%, 26.9%, and 90.6% prevalence rates were confirmed, respectively. This study confirmed the pathogenic E. coli contamination of vegetables to be consumed without cooking. Therefore, to produce agricultural products that do not induce food poisoning and are safe for consumption, it is important to develop a process for killing the pathogenic microorganisms and set up a sanitary environment for effectively managing compost. In addition, it is necessary to establish surveillance systems to monitor the production chain.

A waterborne outbreak involving hepatitis A virus genotype IA at a residential facility in the Republic of Korea in 2015.

BACKGROUND: Hepatitis A virus (HAV), a major cause of acute hepatitis, has had the highest occurrence among group 1 nationally notifiable infectious diseases in Korea since 2010.Recently,the annual increase in the HAV infection rate among young adults has become a public health concern. OBJECTIVES: The aim of this study was to describe an outbreak of acute hepatitis in a residential facility in April 2015 and to identify potential sources of this outbreak. STUDY DESIGN: Sera from all exposed residents were tested for anti-HAV IgM or IgG antibodies by ELISA. Clinical (sera and stool) and environmental samples were screened for the presence of HAV RNA using one-step RT-PCR and nested PCR. The VP3-VP1 regions of HAV were analyzed using the BLAST database and MEGA7 software. RESULTS: Of the 82 persons in the facility, 12 (14.6%, including 10 residents and 2 health care workers) were diagnosed with hepatitis A. Clinical symptoms were evident in 9 individuals, one of whom died, and the remaining four patients were asymptomatic. Traceback investigation revealed that HAV-RNA (genotype IA) was detected in the patients' stools and the groundwater used in the facility. CONCLUSIONS: We described an HAV outbreak in a facility for the disabled due to using a water supply that was mixed with contaminated groundwater. Therefore, HAV vaccination and periodic water inspections in group facilities should be emphasized to prevent HAV infection.