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1.
Immun Inflamm Dis ; 11(2): e783, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36840495

ABSTRACT

BACKGROUND: Sublineage BA.5 of the SARS-CoV-2 Omicron variant rapidly spread and replaced BA.2 in July 2022 in Tokyo. A high viral load can be a possible cause of high transmissibility. METHODS AND RESULTS: The copy numbers of SARS-CoV-2 in nasopharyngeal swab samples obtained from all patients visiting the hospital where this research was conducted were measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Viral genotypes were determined using PCR-based melting curve analysis. Next, whole-genome sequencing was performed using approximately one fifth of the samples to verify the viral genotypes determined using PCR. Then, the copy numbers of the BA.1, BA.2, and BA.5 cases were compared. Contrary to expectations, the copy numbers of the BA.5 cases (median 4.7 × 104 copies/µL, n = 291) were significantly (p = .001) lower than those of BA.2 cases (median 1.1 × 105 copies/µL, n = 184). There was no significant difference (p = .44) between the BA.5 and BA.1 cases (median, 3.3 × 104 copies/µL; n = 215). CONCLUSION: The results presented here suggest that the increased infectivity of BA.5 is not caused by higher viral loads, but presumably by other factors such as increased affinity to human cell receptors or immune escape due to its L452R mutation.


Subject(s)
COVID-19 , Humans , SARS-CoV-2 , Viral Load , Genotype
2.
J Med Virol ; 94(11): 5543-5546, 2022 11.
Article in English | MEDLINE | ID: mdl-35790476

ABSTRACT

Patients infected with the Omicron variant of severe acute respiratory syndrome coronavirus 2 has increased worldwide since the beginning of 2022 and the variant has spread more rapidly than the Delta variant, which spread in the summer of 2021. It is important to clarify the cause of the strong transmissibility of the Omicron variant to control its spread. In 694 patients with coronavirus disease 2019, the copy numbers of virus in nasopharyngeal swab-soaked samples and the viral genotypes were examined using quantitative polymerase chain reaction (PCR) and PCR-based melting curve analysis, respectively. Whole-genome sequencing was also performed to verify the viral genotyping data. There was no significant difference (p = 0.052) in the copy numbers between the Delta variant cases (median 1.5 × 105 copies/µl, n = 174) and Omicron variant cases (median 1.2 × 105 copies/µl, n = 328). During this study, Omicron BA.1 cases (median 1.1 ×105 copies/µl, n = 275) began to be replaced by BA.2 cases (median 2.3 × 105 copies/µl, n = 53), and there was no significant difference between the two groups (p = 0.33). Our results suggest that increased infectivity of the Omicron variant and its derivative BA.2 is not caused by higher viral loads but by other factors, such as increased affinity to cell receptors or immune escape.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , SARS-CoV-2/genetics , Viral Load
3.
J Med Virol ; 94(4): 1707-1710, 2022 04.
Article in English | MEDLINE | ID: mdl-34825717

ABSTRACT

The rapid spread of the Delta variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) became a serious concern worldwide in summer 2021. We examined the copy number and variant types of all SARS-CoV-2-positive patients who visited our hospital from February to August 2021 using polymerase chain reaction (PCR) tests. Whole genome sequencing was performed for some samples. The R.1 variant (B.1.1.316) was responsible for most infections in March, replacing the previous variant (B.1.1.214); the Alpha (B.1.1.7) variant caused most infections in April and May; and the Delta variant (B.1.617.2) was the most prevalent in July and August. There was no significant difference in the copy numbers among the previous variant cases (n = 29, median 3.0 × 104 copies/µl), R.1 variant cases (n = 28, 2.1 × 105 copies/µl), Alpha variant cases (n = 125, 4.1 × 105 copies/µl), and Delta variant cases (n = 106, 2.4 × 105 copies/µl). Patients with Delta variant infection were significantly younger than those infected with R.1 and the previous variants, possibly because many elderly individuals in Tokyo were vaccinated between May and August. There was no significant difference in mortality among the four groups. Our results suggest that the increased infectivity of Delta variant may be caused by factors other than the higher viral loads. Clarifying these factors is important to control the spread of Delta variant infection.


Subject(s)
COVID-19/epidemiology , COVID-19/virology , SARS-CoV-2/physiology , Viral Load , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Mutation , Polymerase Chain Reaction , RNA, Viral/genetics , SARS-CoV-2/classification , SARS-CoV-2/genetics , Tokyo/epidemiology , Whole Genome Sequencing
4.
J Med Virol ; 93(12): 6833-6836, 2021 12.
Article in English | MEDLINE | ID: mdl-34314050

ABSTRACT

The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, such as B.1.1.7 and B.1.351, has become a crucial issue worldwide. Therefore, we began testing all patients with COVID-19 for the N501Y and E484K mutations by using polymerase chain reaction (PCR)-based methods. Nasopharyngeal swab samples from 108 patients who visited our hospital between February and April 2021 were analyzed. The samples were analyzed using reverse transcription-PCR with melting curve analysis to detect the N501Y and E484K mutations. A part of the samples was also subjected to whole-genome sequencing (WGS). Clinical parameters such as mortality and admission to the intensive care unit were analyzed to examine the association between increased disease severity and the E484K mutation. The ratio of cases showing the 501N + 484K mutation rapidly increased from 8% in February to 46% in March. WGS revealed that the viruses with 501N + 484K mutation are R.1 lineage variants. Evidence of increased disease severity related to the R.1 variants was not found. We found that the R.1 lineage variants rapidly prevailed in Tokyo in March 2021, which suggests the increased transmissibility of R.1 variants, while they showed no increased severity.


Subject(s)
COVID-19/epidemiology , COVID-19/virology , SARS-CoV-2/genetics , Aged , Female , Humans , Male , Mutation/genetics , Spike Glycoprotein, Coronavirus/genetics , Tokyo/epidemiology , Whole Genome Sequencing/methods
5.
Cell Tissue Res ; 380(3): 527-537, 2020 Jun.
Article in English | MEDLINE | ID: mdl-31975032

ABSTRACT

Cell migration is essential for many physiological and pathological processes, including embryonic development, wound healing, immune response and cancer metastasis. Inka2 transcripts are observed in migrating cells during embryonic development, suggesting the involvement of inka2 in cell migration. However, its precise role remains unclear. Here, we found that inka2 controlled focal adhesion dynamics and cell migration, likely by regulating protein phosphatase-2A (PP2A) function. A scratch assay revealed that inka2 shRNA-transfected NIH3T3 cells showed rapid wound closure, indicating an inhibitory effect by inka2 on cell migration. Live-cell imaging of NIH3T3 cells expressing EGFP-paxillin using total internal reflection fluorescence microscopy revealed that inka2 knockdown increased the turnover rate of focal adhesions. Given that PP2A, which consists of catalytic (C), regulatory (B) and scaffolding (A) subunits, is known to regulate focal adhesions, we examined the inka2-PP2A interaction. Immunoprecipitation revealed an association between inka2 and the PP2A C subunit. Binding of Inka2 to the C subunit prevented the association between the A and C subunits, suggesting that inka2 can inhibit PP2A function. Furthermore, both inka2 expression and PP2A inhibition decreased focal adhesion kinase-paxillin interaction, resulting in reduced formation of focal adhesions. We assessed the effect of pharmacological PP2A inhibition on the inka2 knockdown-induced increase in cell migration speed and found that treatment with a PP2A inhibitor negated the accelerated migration of inka2 knockdown cells. These results suggest that inka2 knockdown exerts its effects through PP2A-dependent regulation of focal adhesions. Our findings contribute to a better understanding of the molecular mechanisms underlying cell migration.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Cell Movement , Intracellular Signaling Peptides and Proteins/metabolism , Protein Phosphatase 2/metabolism , Animals , Focal Adhesions , Gene Knockdown Techniques , HEK293 Cells , Humans , Mice , Mice, Inbred ICR , NIH 3T3 Cells
6.
Biosci Biotechnol Biochem ; 80(12): 2401-2411, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27573376

ABSTRACT

Various commercial tomato juices with different flavors are available at markets worldwide. To clarify the marker compounds related to the flavor characteristics of tomato juice, we analyzed 15 pure commercial tomato juices by a combination of volatile profiling and sensory evaluation. The correlations among volatiles and the relationship between volatiles and sensory descriptors were elucidated by multivariate analyses. Consequently, the tomato juices made from fresh market tomatoes (including the popular Japanese tomato variety "Momotaro") were clearly separated from other juices made from processing tomatoes, by both the volatile composition and sensory profiles. cis-3-Hexenol, hexanal, and apocarotenoids negatively contributed to the juices from fresh market tomatoes, whereas Strecker aldehydes and furfural showed positive contributions to the juices. Accordingly, the sensory characteristics of juices from fresh market tomatoes were related to cooked and fruity flavors but not to green or fresh notes.


Subject(s)
Food Handling , Fruit and Vegetable Juices/analysis , Fruit/chemistry , Solanum lycopersicum/chemistry , Taste/drug effects , Volatile Organic Compounds/analysis , Volatile Organic Compounds/pharmacology , Humans , Volatile Organic Compounds/chemistry
7.
Gene Expr Patterns ; 19(1-2): 83-97, 2015.
Article in English | MEDLINE | ID: mdl-26292052

ABSTRACT

Dynamic rearrangement of the actin cytoskeleton impacts many cellular characteristics in both the developing and adult central nervous systems (CNS), including the migration and adhesion of highly motile neural progenitor cells, axon guidance of immature neurons, and reconstruction of synaptic structures in the adult brain. Inka1, a known regulator of actin cytoskeleton reconstruction, is predominantly expressed by the neural crest cell lineage and regulates the migration and differentiation of these cells. In the present study, we identified a novel gene, designated as inka2, which is related to inka1. Inka2/fam212b is an evolutionarily conserved gene found in different vertebrate species and constitutes a novel gene family together with inka1. Northern blot analysis showed that inka2 mRNA was highly enriched in the nervous system. The spatiotemporal propagation cell profiles of those cells that expressed inka2 transcripts were compatible with those of Olig2-positive oligodendrocyte progenitor cells, which originate in the ventral ventricular zone during embryogenesis. Intense expression of inka2 was also noted in the proliferative neuronal progenitors in the developing cerebellum. On the other hand, immature newborn neurons in the embryonic brain showed no expression of inka2, except for the cells residing in the marginal zone of the embryonic telencephalon, which is known to contain transient cells including the non-subplate pioneer neurons and Cajal-Retzius cells. As brain development proceeds during the postnatal stage, inka2 expression emerged in some populations of immature neurons, including the neocortical pyramidal neurons, hippocampal pyramidal neurons, and granule cells migrating in the cerebellar cortex. In the adult brain, the expression of inka2 was interestingly confined in terminally differentiated neurons in the restricted forebrain regions. Taken together, as a novel regulator of actin cytoskeletons in the CNS, inka2 may be involved in multiple actin-driven processes, including cell migration and establishment of neuronal polarity.


Subject(s)
Intracellular Signaling Peptides and Proteins/genetics , Nerve Tissue Proteins/genetics , Animals , Cell Differentiation/physiology , Cell Movement/physiology , Gene Expression Profiling , Gene Expression Regulation, Developmental , Hippocampus/cytology , Intracellular Signaling Peptides and Proteins/biosynthesis , Male , Mice , Mice, Inbred ICR , Neocortex/cytology , Neural Stem Cells/metabolism , Neurons/metabolism , Oligodendroglia/metabolism , Prosencephalon/cytology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Stem Cells/metabolism
8.
Chemosphere ; 93(6): 1222-9, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23916213

ABSTRACT

Despite the enormous cost of radiation decontamination, there has been almost no quantitative discussion on how much it would reduce the long-term external radiation exposure in the Evacuation Zone and Planned Evacuation Zone (restricted zone) in Fukushima. The aim of this study is to assess the effectiveness of decontamination and return options and to identify important parameters for estimating the long-term cumulated effective dose (CED) during 15, 30 and 70 year period using data on land-use, population and decontamination in the restricted zone (about 1100 km(2)) in Fukushima. Decontamination of the land is assumed to have a certain efficacy in terms of the reduction of CED. The EeCC (external exposure conversion coefficient) is the parameter having the greatest effect on the percentage of area having CED during the 30 years above 100 m Sv after decontamination, ranging from 13% (EeCC=0.2) to 55% (EeCC=0.6). Therefore, we recommend a detailed investigation of the EeCC in Japan.


Subject(s)
Decontamination , Fukushima Nuclear Accident , Geographic Information Systems , Radiation Dosage , Radiation Monitoring , Radioactive Pollutants/analysis , Humans , Japan , Nuclear Power Plants
9.
Gan To Kagaku Ryoho ; 37(9): 1753-7, 2010 Sep.
Article in Japanese | MEDLINE | ID: mdl-20841940

ABSTRACT

The recent guidelines of the Japanese Society of Hospital Pharmacists on the antitumor drug preparation have recommended the use of closed systems such as the PhaSeal® system for preventing cytotoxicity in health care workers involved in the preparation of these drugs. The PhaSeal® system and Clave® Oncology system were evaluated using a practical training kit for the preparation of antitumor drugs. The two systems were compared in terms of handling time, satisfaction as to availability, leakage of drugs from the connections in the system and area of drug spillage because improvements in convenience or lower cost system were available. With the closed systems, the average handling time increased by 10∼20%. The area of drug spillage did not significantly decrease. Leakage of drugs from the system was detected for all samples prepared with the Clave® Oncology system, and for some samples prepared with the PhaSeal® system. In terms of availability, the PhaSeal® system was better than the Clave® Oncology system. In conclusion, to decrease the exposure of health care workers to antitumor drugs during their preparation in a closed system, it is important to evaluate the handling time, operability, robustness with regard to drug leakage and spillage, and proficiency in handling of the closed system.


Subject(s)
Antineoplastic Agents/analysis , Environmental Monitoring/methods , Antineoplastic Agents/poisoning , Practice Guidelines as Topic
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