Relationship between Internet use and out-of-home activities during the first wave of the COVID-19 outbreak in Japan.

Following the first wave of the COVID-19 outbreak, the Japanese government announced the declaration of a state of emergency in April 2020, which aimed to decrease contact between people and requested that residents refrain from outings. Even in the absence of penalties, outings decreased under the declaration. We are interested in how outings declined and studied the substitution relationship between Internet use and outings. A web-based survey was conducted to collect data on Internet use and outings in a retrospective manner. The period covered by our data is from mid-February to mid-May 2020. Multilevel analysis and binomial logistic regression analysis were performed to examine the relationship between Internet use and outings. The results clearly show that Internet use replaced outings. In particular, Internet use for socializing, exercise, and leisure/entertainment had a strong substitution relationship with outings. Internet use for socializing and leisure/entertainment was also associated with refraining from visiting restaurants. In contrast, there was a weak substitution relationship between Internet use for daily shopping and outings. Although telework tends to be an accepted focus of Internet use under the COVID-19 outbreak, it should not be overlooked that other uses of the Internet, such as for leisure/entertainment, also supported the decline in outings.

Who is staying home and who is not? Demographic, socioeconomic, and geographic differences in time spent outside the home during the COVID-19 outbreak in Japan.

Studies have reported that many people changed their going-out behavior in response to the declaration of a state of emergency related to the coronavirus disease 2019 (COVID-19) in Japan. However, individual attributes of those who tended to stay home have not been examined. Therefore, this study examined the demographic, socioeconomic, and geographic characteristics of people who refrained from going out both before and after a state of emergency was declared. Using data from a nationwide online survey, this study retrospectively investigated the relative amount of time spent outside the home between mid-February and mid-May 2020. Multilevel linear regression analysis was performed to examine the association of time outside with demographic, socioeconomic, and geographic characteristics, and with the anxiety related to going out, in each period. Overall, respondents significantly reduced their time spent outside during the study period, especially after a state of emergency was declared. Those who were young, female, living with two or more people, had lower income, were not working, used public transportation, had chronic disease, and lived in large metropolitan areas were more likely to reduce time outside during a part of the study period. However, no significant differences were observed for occupational class, education, and neighborhood population density. Thus, the results showed a reduction in time outside during the COVID-19 outbreak and the existence of demographic, socioeconomic, and geographic differences in going-out behavior. Socioeconomic disparities and neighborhood differences in going-out behavior, and their influence on health should be continuously monitored.

Metabolic changes in Arabidopsis thaliana expressing the feedback-resistant anthranilate synthase alpha subunit gene OASA1D.

Anthranilate synthase (AS) is a key enzyme in tryptophan (Trp) biosynthesis. Metabolic changes in transgenic Arabidopsis plants expressing the feedback-resistant anthranilate synthase alpha subunit gene OASA1D were investigated with respect to Trp synthesis and effects on secondary metabolism. The Trp content varied depending on the transgenic line, with some lines showing an approximately 200-fold increase. The levels of AS activity in crude extracts from the transgenic lines were comparable to those in the wild type. On the other hand, the enzyme prepared from the lines accumulating high levels of Trp showed a relaxed feedback sensitivity. The AS activity, determined in the presence of 50 microM L-Trp, correlated well with the amount of free Trp in the transgenic lines, indicating the important role of feedback inhibition in control of Trp pool size. In Arabidopsis, Trp is a precursor of multiple secondary metabolites, including indole glucosinolates and camalexin. The amount of indol-3-ylmethyl glucosinolate (I3 M) in rosette leaves of the high-Trp accumulating lines was 1.5- to 2.1-fold greater than that in wild type. The treatment of the leaves with jasmonic acid resulted in a more pronounced accumulation of I3 M in the high-Trp accumulating lines than in wild type. The induction of camalexin formation after the inoculation of Alternaria brassicicola was not affected by the accumulation of a large amount of Trp. The accumulation of constitutive phenylpropanoids and flavonoids was suppressed in high-Trp accumulating lines, while the amounts of Phe and Tyr increased, thereby indicating an interaction between the Trp branch and the Phe and Tyr branch in the shikimate pathway.

Characterization of the class IV homeodomain-Leucine Zipper gene family in Arabidopsis.

The Arabidopsis (Arabidopsis thaliana) genome contains 16 genes belonging to the class IV homeodomain-Leucine zipper gene family. These include GLABRA2, ANTHOCYANINLESS2, FWA, ARABIDOPSIS THALIANA MERISTEM LAYER1 (ATML1), and PROTODERMAL FACTOR2 (PDF2). Our previous study revealed that atml1 pdf2 double mutants have severe defects in the shoot epidermal cell differentiation. Here, we have characterized additional members of this gene family, which we designated HOMEODOMAIN GLABROUS1 (HDG1) through HDG12. Analyses of transgenic Arabidopsis plants carrying the gene-specific promoter fused to the bacterial beta-glucuronidase reporter gene revealed that some of the promoters have high activities in the epidermal layer of the shoot apical meristem and developing shoot organs, while others are temporarily active during reproductive organ development. Expression profiles of highly conserved paralogous gene pairs within the family were found to be not necessarily overlapping. Analyses of T-DNA insertion mutants of these HDG genes revealed that all mutants except hdg11 alleles exhibit no abnormal phenotypes. hdg11 mutants show excess branching of the trichome. This phenotype is enhanced in hdg11 hdg12 double mutants. Double mutants were constructed for other paralogous gene pairs and genes within the same subfamily. However, novel phenotypes were observed only for hdg3 atml1 and hdg3 pdf2 mutants that both exhibited defects in cotyledon development. These observations suggest that some of the class IV homeodomain-Leucine zipper members act redundantly with other members of the family during various aspects of cell differentiation. DNA-binding sites were determined for two of the family members using polymerase chain reaction-assisted DNA selection from random oligonucleotides with their recombinant proteins. The binding sites were found to be similar to those previously identified for ATML1 and PDF2, which correspond to the pseudopalindromic sequence 5'-GCATTAAATGC-3' as the preferential binding site.

Overexpression of constitutive differential growth 1 gene, which encodes a RLCKVII-subfamily protein kinase, causes abnormal differential and elongation growth after organ differentiation in Arabidopsis.

To better understand genetic regulation of differential growth of plant organs, a dominant and semidwarf mutant, constitutive differential growth 1-Dominant (cdg1-D), was isolated utilizing the technique of activation tagging. cdg1-D showed pleiotropic phenotype including dwarfism, exaggerated leaf epinasty, and twisted or spiral growth in hypocotyl, inflorescence stem, and petiole. Hypocotyls of cdg1-D were longer than those of wild type under light conditions. The phenotype was caused by activation tagging of CDG1 gene that encodes a receptor-like cytoplasmic kinase of RLCKVII subfamily. When treated with high concentrations of brassinolide, light-grown wild-type seedlings showed long hypocotyls and strong leaf epinasty as observed in cdg1-D seedlings. Treatment of cdg1-D with brassinazole, a specific inhibitor of brassinosteroid (BR) biosynthesis, did not rescue the mutant phenotype. Gene expression of CONSTITUTIVE PHOTOMORPHOGENESIS AND DWARFISM involved in BR biosynthesis and phyB ACTIVATION-TAGGED SUPPRESSOR1 that inactivates BR was repressed and induced, respectively, in cdg1-D plants, suggesting constitutive activation of BR signaling in the mutant. CDG1 was expressed at a very low level in all the organs of the wild type tested. We isolated two independent intragenic suppressors of cdg1-D. However, they showed normal morphology and responded to BR in a similar manner to wild type. Taken together, CDG1 gene may interfere with signal transduction of BR when overexpressed, but is not an essential factor for it in the wild type.

A methyl viologen-resistant mutant of Arabidopsis, which is allelic to ozone-sensitive rcd1, is tolerant to supplemental ultraviolet-B irradiation.

To better understand the role of active oxygen species (AOS) in acquired resistance to increased levels of ultraviolet (UV)-B irradiation in plants, we isolated an Arabidopsis mutant that is resistant to methyl viologen, and its sensitivity to UV-B was investigated. A complementation test revealed that the obtained mutant was allelic to the ozone-sensitive radical-induced cell death1-1 (rcd1-1). Therefore, this mutant was named rcd1-2. rcd1-2 was recessive and nearly 4-fold more resistant to methyl viologen than wild type. It exhibited a higher tolerance to short-term UV-B supplementation treatments than the wild type: UV-B-induced formation of cyclobutane pyrimidine dimers was reduced by one-half after 24 h of exposure; the decrease in quantum yield of photosystem II was also diminished by 40% after 12 h of treatment. Furthermore, rcd1-2 was tolerant to freezing. Steady-state mRNA levels of plastidic Cu/Zn superoxide dismutase and stromal ascorbate peroxidase were higher in rcd1-2 than in wild type, and the mRNA level of the latter enzyme was enhanced by UV-B exposure more effectively in rcd1-2. UV-B-absorbing compounds were more accumulated in rcd1-2 than in wild type after UV-B exposure for 24 h. These findings suggest that rcd1-2 methyl viologen resistance is due to the enhanced activities of the AOS-scavenging enzymes in chloroplasts and that the acquired tolerance to the short-term UV-B exposure results from a higher accumulation of sunscreen pigments. rcd1 appears to be a mutant that constitutively shows stress responses, leading to accumulation of more pigments and AOS-scavenging enzymes without any stresses.

Arabidopsis NDK1 is a component of ROS signaling by interacting with three catalases.

Plants sense various environmental stimuli and have specific signaling pathways to respond to these cues. We focused on light responsive components and found that NDKs were phosphorylated specifically after red light irradiation in Pisum sativum [Tanaka et al. (1998) J. Photochem. Photobiol. B 45: 113] and after blue light irradiation in Neurospora crassa [Oda and Hasunuma (1997) Mol. Gen. Genet. 256: 593, Ogura et al. (2001) J. Biol. Chem. 276: 21228]. We performed yeast two-hybrid screening using AtNDK1, the counterpart of NDK-P1 (Pisum sativum NDK1) in Arabidopsis, as bait, and isolated catalase3 (AtCat3). Interactions between AtNDK1-AtCAT1 and AtNDK1-AtCAT2 were also detected with the two-hybrid system. Non-denaturing two-dimensional gel electrophoresis of crude extracts from plants revealed that catalase and NDK activities co-migrated in the same area of the gel. Transgenic plants expressing AtNDK1 under control of the CaMV 35S promoter exhibited tolerance to paraquat and high ability to eliminate exogenous H2O2. These results indicate that AtNDK1 has a role in ROS response.

Putative functions of nucleoside diphosphate kinase in plants and fungi.

The putative functions of NDP (nucleoside diaphosphate) kinases from various organisms focusing to fungi and plants are described. The biochemical reactions catalyzed by NDP kinase are as follows. (i) Phosphotransferring activity from mainly ATP to cognate NDPs generating nucleoside triphosphates (NTPs). (ii) Autophosphorylation activity from ATP and GTP. (iii) Protein kinase (phosphotransferring) activity phosphorylating such as myelin basic protein. NDP kinase could function to provide NTPs as a housekeeping enzyme. However, recent works proved possible functions of the NDP kinases in the processes of signal transduction in various organisms, as described below. 1) By use of the extracts of the mycelia of a filamentous fungus Neurospora crassa blue-light irradiation could increase the phosphorylation of a 15-kDa protein, which was purified and identified to be NDP kinase (NDK-1). By use of the etiolated seedlings of Pisum sativum cv Alaska and Oryza sativa red-light irradiation of intact plants increased the phosphorylation of NDP kinase. However, successive irradiation by red-far-red reversed the reaction, indicating that phytochrome-mediated light signals are transduced to the phosphorylation of NDP kinase. 2) NDP kinase localizing in mitochondria is encoded by nuclear genome and different from those localized in cytoplasm. NDP kinase in mitochondria formed a complex with succinyl CoA synthetase. 3) In Spinicia oleraceae two different NDP kinases were detected in the chloroplast, and in Pisum sativum two forms of NDP kinase originated from single species of mRNA could be detected in the choloroplast. However, the function of NDP kinases in the choloroplast is not yet known. 4) In Neurospora crassa a Pro72His mutation in NDP kinase (ndk-1Pro72His) deficient in the autophosphorylation and protein kinase activity resulted in lacking the light-induced polarity of perithecia. In wild-type directional light irradiation parallel to the solid medium resulted in the formation of the perithecial beak at the top of perithecia, which was designated as "light-induced polarity of perithecia." In wild-type in darkness the beak was formed at random places on perithecia, and in ndkPro72His mutant the perithecial beak was formed at random places even under directional light illumination. The introduction of genomic DNA and cDNA for ndk-1 demonstrated that the wild-type DNAs suppressed the mutant phenotype. With all these results except for the demonstration in Neurospora, most of the phenomena are elusive and should be solved in the molecular levels concerning with NDP kinases.