Objectives The aim was to determine whether various clinical specimens obtained from COVID-19 patients contain the infectious virus. Methods To demonstrate whether various clinical specimens contain the viable virus, we collected naso/oropharyngeal swabs and saliva, urine and stool samples from five COVID-19 patients and performed a quantitative polymerase chain reaction (qPCR) to assess viral load. Specimens positive with qPCR were subjected to virus isolation in Vero cells. We also used urine and stool samples to intranasally inoculate ferrets and evaluated the virus titres in nasal washes on 2, 4, 6 and 8 days post infection. Results SARS-CoV-2 RNA was detected in all naso/oropharyngeal swabs and saliva, urine and stool samples collected between days 8 and 30 of the clinical course. Notably, viral loads in urine, saliva and stool samples were almost equal to or higher than those in naso/oropharyngeal swabs (urine 1.08 ± 0.16–2.09 ± 0.85 log 10 copies/mL, saliva 1.07 ± 0.34–1.65 ± 0.46 log 10 copies/mL, stool 1.17 ± 0.32 log 10 copies/mL, naso/oropharyngeal swabs 1.18 ± 0.12–1.34 ± 0.30 log 10 copies/mL). Further, viable SARS-CoV-2 was isolated from naso/oropharyngeal swabs and saliva of COVID-19 patients, as well as nasal washes of ferrets inoculated with patient urine or stool. Discussion Viable SARS-CoV-2 was demonstrated in saliva, urine and stool samples from COVID-19 patients up to days 11–15 of the clinical course. This result suggests that viable SARS-CoV-2 can be secreted in various clinical samples and respiratory specimens.
Plant growth promoting rhizobacteria (PGPR) are known to confer disease resistance to plants. Bacillus sp. JS demonstrated antifungal activities against five fungal pathogens in in vitro assays. To verify whether the volatiles of Bacillus sp. JS confer disease resistance, tobacco leaves pre-treated with the volatiles were damaged by the fungal pathogen, Rhizoctonia solani and oomycete Phytophthora nicotianae. Pre-treated tobacco leaves had smaller lesion than the control plant leaves. In pathogenesis-related (PR) gene expression analysis, volatiles of Bacillus sp. JS caused the up-regulation of PR-2 encoding β-1,3-glucanase and acidic PR-3 encoding chitinase. Expression of acidic PR-4 encoding chitinase and acidic PR-9 encoding peroxidase increased gradually after exposure of the volatiles to Bacillus sp. JS. Basic PR-14 encoding lipid transfer protein was also increased. However, PR-1 genes, as markers of salicylic acid (SA) induced resistance, were not expressed. These results suggested that the volatiles of Bacillus sp. JS confer disease resistance against fungal and oomycete pathogens through PR genes expression.
New multiple-stress related gene isolated from sweet potato and designated it as MusI (multiple stress responsible gene I). Sequence analysis revealed that its full length cDNA was 998 bp long and included a 717 bp open reading frame encoding for 238 amino acids. Comparison of its cDNA and genomic DNA sequence showed that 3 exons were divided by 2 introns in its ORF region. Its deduced amino acid sequence contained a conserved rubber elongation factor (REF) domain and showed high homology with many stress-related proteins. Therefore, it was named MuSI (multiple stress responsible gene I). Southern hybridization analysis indicated that the MuSI gene may belong to a multi-gene family. Expression pattern of the MuSI gene showed that it was differently expressed among roots, stems, leaves, and flowers of a sweet potato, and its expression level was especially high in flowers andThe first two authors, SGS and JSK, contributed equally. white fibrous roots. Its expression was also highly induced by various stress signals including dehydration, high salt, heavy metal, oxidation, and plant hormones. Stress tolerance experiment using transgenic plants overexpressing the MuSI gene showed that all independent transgenic tobacco lines have enhanced tolerance to high temperature stress. Among them, transgenic line 6 particularly showed tolerance to salt, heavy metal, and osmotic stress as well. These results suggest that the MuSI gene functions as a positive regulator of various stress responses and may be useful in improving stress tolerance of transgenic plants.
Background: Opioid analgesics are generally used to combat the pain associated with cancerous conditions. These agents not only inhibit respiratory function and cause constipation, but also induce other significant side effects such as addiction and tolerance, all of which further contribute to a reduced quality of life for cancer patients. Thus, in the present study, the effects of electroacupuncture treatment (EA) on mechanical allodynia were examined in a cancer pain mouse model.
Volatile and nonvolatile compounds emitted from the plant growth-promoting rhizobacterium Bacillus sp. strain JS enhance the growth of tobacco and lettuce. Here, we report the high-quality genome sequence of this bacterium. Its 4.1-Mb genome reveals a number of genes whose products are possibly involved in promotion of plant growth or antibiosis.
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