ABSTRACT
Since the outbreak of the COVID-19 global pandemic in 2019, monitoring COVID-19 infection status and trend through wastewater, known as wastewater-based epidemiology (WBE), has been widely used in many countries and regions. WBE consists of five steps:wastewater sample collection, viral concentration, viral nucleic acid extraction, quantification of virus using quantitative RT-PCR, and dissemination of the wastewater surveillance results. This method could be used for early warning of COVID-19 outbreak in a population, monitoring COVID-19 distributions and epidemic trend, prediction of COVID-19 prevalence rate, understanding of temporal trend of SARS-COV-2 variants, and simultaneous surveillance of multiple pathogens. WBE and clinical surveillance can be used concurrently and the former is a good complement to the latter.
ABSTRACT
Little is known about the effectiveness of disinfectants against human noroviruses [NoV] partially because human NoV cannot be routinely cultured in laboratory. The objective of this study was to develop a NoV monoclonal antibody-conjugated immunomagnetic separation [IMS] procedure combined with real-time reverse transcription polymerase chain reaction [RT-qPCR] assays to study the in vitro efficacy of disinfectants against human NoV. Monoclonal antibodies against Norwalk virus [NV, GI.1] and NoV GII.4 were produced using unique NoV capsid proteins, and the antibodies were conjugated to magnetic Dynalbeads. The immunomagnetic beads were used to simultaneously capture intact NoV in samples and effectively remove PCR inhibitors. We examined the efficacy of ethanol, sodium hypochlorite, nine commercially available disinfectants, and one prototype disinfectant using the IMS/RT-qPCR. The sensitivity of this procedure was approximately 100 virus particles for both the NV and GII.4 viruses. The average log reductions in in vitro activities varied between disinfectants. The prototype disinfectant produced an average 3.19-log reduction in NV and a 1.38-log reduction in GII.4. The prototype disinfectant is promising of inactivating NoV. This method can be used to evaluate in vitro activity of disinfectants against human NoV. The IMS/RT-qPCR method is promising as an effective method to remove PCR inhibitors in disinfectants and enable the evaluation of the efficacy of disinfectants
Subject(s)
Norovirus/drug effects , Immunomagnetic Separation , Reverse Transcriptase Polymerase Chain Reaction , Real-Time Polymerase Chain ReactionABSTRACT
Through studying the process of glycerol fermentation to 1, 3-propanediol(1, 3-PD) by Klebsiella pneumoniae, it was found that the cell growth and product (or by-product) production were under salt stress. Cell growth and product formation kept high rate at low salt concentration. High salt concentration led to low growth of cells, final concentration of 1, 3-PD and conversion from glycerol to 1, 3-PD, and, 1, 3-propanediol oxidoreductase activity decreased. When the salt concentration in 5 m3 bioreactor was controlled under appropriate manner, the concentration of 1, 3-PD production was markedly enhanced. The final 1, 3-PD concentration ,the conversion of glycerol to 1, 3-PD and productivity were 64 g/L, 61% and 2.1 g/(L x h).
Subject(s)
Alcohol Dehydrogenase , Alcohol Oxidoreductases , Metabolism , Culture Media , Culture Techniques , Fermentation , Glycerol , Metabolism , Klebsiella pneumoniae , Metabolism , Physiology , Propylene Glycols , Metabolism , Sodium Chloride , Pharmacology , Stress, PhysiologicalABSTRACT
cDNA microarray was used to compare the gone expression profiles of multiple myeloma cell line RPMI8226 24 h before and after treatment with arsenic trioxide. Two eDNA probes were prepared by mRNA reverse transcription of both arsenic trioxide-treated and untreated RPMI8226 cells. The probes were labeled with Cy3 and Cy5 fluorescence dyes separately, hybridized with cDNA microarray representing 4096 different human genes, and scanned for fluorescence intensity. The differences in gene expression were calculated on the basis of the ratios of signal intensity of treated and untreated samples. The up- and down-regulated genes were screened through the analysis of gene expression ratios. The results showed that 273 genes were differentially altered at mRNA level, 121 genes were up-regulated and 152 were down-regulated. It is concluded that the treatment with arsenic trioxide can induce a variety of gene changes in RPMI8226 cell line. Many genes may be involved in the pathogenesis of multiple myeloma. ALK-1 and TXNIP genes may play an impor- tant role in the apoptosis and partial differentiation of RPMI8226 cells.
ABSTRACT
cDNA microarray was used to compare the gene expression profiles of multiple myeloma cell line RPMI8226 24 h before and after treatment with arsenic trioxide. Two cDNA probes were prepared by mRNA reverse transcription of both arsenic trioxide-treated and untreated RPMI8226 cells. The probes were labeled with Cy3 and Cy5 fluorescence dyes separately, hybridized with cDNA microarray representing 4096 different human genes, and scanned for fluorescence intensity. The differences in gene expression were calculated on the basis of the ratios of signal intensity of treated and untreated samples. The up-and down-regulated genes were screened through the analysis of gene expression ratios. The results showed that 273 genes were differentially altered at mRNA level, 121 genes were up-regulated and 152 were down-regulated. It is concluded that the treatment with arsenic trioxide can induce a variety of gene changes in RPMI8226 cell line. Many genes may be involved in the pathogenesis of multiple myeloma. ALK-1 and TXNIP genes may play an important role in the apoptosis and partial differentiation of RPMI8226 cells.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Arsenicals/pharmacology , Gene Expression Profiling , Multiple Myeloma/genetics , Multiple Myeloma/pathology , Oxides/pharmacology , Tumor Cells, CulturedABSTRACT
OBJECTIVE: To compare the changes of gene expression profiles of multiple myeloma cell line RPMI 8226 before and after 24-hour intervention of arsenic trioxide. METHODS: The responses of the RPMI 8226 cells to arsenic trioxide were determined with cDNA microarray which included 4,096 different human genes. RESULTS: Of these 4,096 genes, the expressions of 273 genes were altered significantly at mRNA level. The expressions of 121 genes were up-regulated while the expressions of 152 genes were down-regulated. CONCLUSION: The effect of arsenic trioxide on RPMI 8226 cells is related to changing the expression levels of a number of genes. ZFYVE16, ALK1 and TXNIP genes may play important roles in apoptosis and differentiation of RPMI 8226 cells.