Tracking community infection dynamics of COVID-19 by monitoring SARS-CoV-2 RNA in wastewater, counting positive reactions by qPCR.

Wastewater-based epidemiology has proved useful for monitoring the COVID-19 infection dynamics in communities. However, in regions of low prevalence, low concentrations of SARS-CoV-2 RNA in wastewater make this difficult. Here, we used real-time reverse-transcription PCR (RT-qPCR) to monitor SARS-CoV-2 RNA in wastewater from October 2020 to December 2022 during the third, fourth, fifth, sixth, seventh, and eighth waves of the COVID-19 outbreak in Japan. Viral RNA was below the limit of detection in all samples during the third and fourth waves. However, by counting the number of positive replicates in qPCR of each sample, we found that the positive ratio to all replicates in wastewater was significantly correlated with the number of clinically confirmed cases by the date of symptom onset during the third, fourth, and fifth waves. Time-step analysis indicated that, for 2 days either side of symptom onset, COVID-19 patients excreted in their feces large amounts of virus that wastewater surveillance could detect. We also demonstrated that the viral genome copy number in wastewater, as estimated from the positive ratio of SARSA-CoV-2 RNA, was correlated with the number of clinically confirmed cases. The positive count method is thus useful for tracing COVID-19 dynamics in regions of low prevalence.

Three embryonic myosin heavy chain genes encoding different motor domain structures from common carp show distinct expression patterns in cranial muscles.

Three embryonic myosin heavy chain (MYH) genes >> (MYHs) including MYH(emb1), MYH(emb2) and MYH(emb3) and encoding a C-terminal part of MYH were previously cloned and demonstrated to be expressed transiently in this order during development of common carp Cyprinus carpio embryos. The present study determined the full-length cDNA nucleotide sequences encoding the motor domain of the three MYHs, suggesting the implication of loop 1 and loop 2 sequences for the differences in the motor functions. Phylogenetic analysis based on the full-length amino acid sequences showed that MYH(emb1) and MYH(emb2) both belong to the fast types, though clearly differ from fast-type MYHs expressed in adult fast muscle previously reported. In contrast, MYH(emb3) was in a clade containing slow/cardiac type. Whole-mount immunostaining and in situ hybridization showed that the transcripts of the three embryonic MYHs are localized in the same or different cranial muscles of common carp larvae, suggesting that the three MYHs function cooperatively or individually in various cranial muscles.

Sex differences in beliefs about cues to deception.

Sex differences in beliefs among Japanese students about cues to deception were explored. 171 participants (91 women, 80 men) read a scenario in which a protagonist caused a fatal traffic accident and told a lie to avoid responsibility. Then participants rated how the protagonist's behaviors would change when lying. Women participants believed significantly more than men that a liar shows body cues (e.g., body touching, biting lips) associated with anxiety, and that a liar has unsuccessful impression management (e.g., fewer smiles, fewer facial expressions). Furthermore, the women's scores also indicated that a liar would increase the amount of information (e.g., longer response length, gestures) and show more nonfluent speech (e.g., speech disturbances, inconsistency of speech contents).

Understanding the concept of a 'good death' in Japan: differences in the views of doctors, palliative and non-palliative ward nurses.

AIM: Achieving a 'good death' is the ultimate goal for medical staff caring for terminally ill cancer patients, but the exact definition of a 'good death' is subject to interpretation. This exploratory study investigates the differences in how Japanese cancer care staff perceive a 'good death' by using factor analysis. METHOD: Participants included doctors, palliative and non-palliative ward nurses working in Miyagi Cancer Center, Natori, Japan. The various components of a 'good death' for cancer patients have been identified in a previous Japanese study. In the present study, respondents were asked to state the percentages of patients for which the component had been achieved, the extent to which the respondent contributed to achieving the component, and the concrete means they would use to ensure that patients were free from psychological distress and prepared for dying. RESULTS: Medical staff had largely similar views on the percentage of patients for which a component of 'good death' was achieved. All the achievement ratings determined in the present study were considerably lower than the necessity ratings given by non-medical staff in a previous study. There were differences among medical staff with respect to their contribution to achieving a component of 'good death' and the concrete means they would use to achieve a 'good death'.

[Retelling harsh days as being happy: its effects on autobiographical memories].

This study examined the effects of biased retelling by having participants retell negative experiences as positive autobiographical memories. Undergraduates in the biased-retelling condition (N = 21) retold their experiences preparing for university entrance examinations as "happy", while undergraduates in the recalling (control) condition (N = 23) again narrated their actual experiences. Then both groups were asked to recall their initial experience. Their initial memories were compared to those after the biased retellings or repeated narrations. In the recall of the autobiographical memories after the biased retellings, the results showed significantly increased positive emotional words and decreased negative emotional words. The emotional values of the central and peripheral concepts of the harsh experiences changed in the direction of "happy" in the biased retelling condition compared to the repeated recalling condition. Furthermore, the changes in the emotional values were more prominent in the central concepts of undergraduates' experiences.

Identification and characterization of mouse type II platelet-derived growth factor receptor alpha transcript.

Recently, we identified a novel human platelet-derived growth factor (PDGF) receptor alpha mRNA (type II) that differs from the known human PDGF receptor alpha mRNA (type I) only in its 5'-untranslated region and is regulated by E2F-1. In this study, we determined the nucleotide sequence of the mouse type II PDGF receptor alpha mRNA and found that the expression of this transcript was regulated by E2F-1 through an E2F-1-responsive sequence located +711/+718 downstream of the transcription start site of the type I transcript. Furthermore, we observed embryo-predominant expression of the type II transcript.

Transcriptional regulation of a new variant of human platelet-derived growth factor receptor alpha transcript by E2F-1.

Platelet-derived growth factors (PDGFs) and their receptors play an important role in cell proliferation, angiogenesis, and differentiation during normal development, and have also been implicated in tumorigenesis. In this study, we identified a novel variant of human PDGF receptor alpha mRNA (type II), which contains the same open reading frame as the known PDGF receptor alpha mRNA (type I) but a different 5'-untranslated region (5'-UTR). The 5'-UTR of the type II transcript was identified as a 363-bp exon located in intron 1 at position +1,210 to +1,572 relative to the transcriptional initiation site of the type I transcript. This type II transcript was expressed in a subset of human cell lines, such as MG-63 and MNNG/HOS cells. Moreover, transcription of the type II, but not the type I, was regulated by E2F-1 through an E2F-1-responsive site located at position +1,086/+1,093 downstream of the transcriptional initiation site of the type I transcript. Furthermore, epigenetic modulation might be involved in the expression of the type II transcript. Our findings provide new insights into the regulatory mechanism of PDGF receptor alpha transcription in normal and tumor cells.

Molecular cloning and mRNA expression analysis of carp embryonic, slow and cardiac myosin heavy chain isoforms.

Three embryonic class II myosin heavy chains (MYHs) were cloned from the common carp (Cyprinus carpio L.), MYHemb1, MYHemb2 and MYHemb3. MYH DNA clones were also isolated from the slow muscle of adult carp acclimated to 10 degrees C (MYHS10) and 30 degrees C (MYHS30). Phylogenetic analysis demonstrated that MYHemb1 and MYHemb2 belonged to the fast skeletal muscle MYH clade. By contrast, the sequence of MYHemb3 was similar to the adult slow muscle isoforms, MYHS10 and MYHS30. MYHemb1 and MYHemb2 transcripts were first detected by northern blot analysis in embryos 61 h post-fertilization (h.p.f.) at the heartbeat stage, with peak expression occurring in 1-month-old juveniles. MYHemb1 continued to be expressed at low levels in 7-month-old juveniles when MYHemb2 was not detectable. MYHemb3 transcripts appeared at almost the same stage as MYHemb1 transcripts did (61 h.p.f.), and these genes showed a similar pattern of expression. Whole mount in situ hybridization analysis revealed that the transcripts of MYHemb1 and MYHemb2 were expressed in the inner part of myotome, whereas MYHemb3 was expressed in the superficial compartment. MYHS10 and MYHS30 mRNAs were first detected at hatching. In adult stages, the expression of slow muscle MYH mRNAs was dependent on acclimation temperature. MYHS10 mRNA was expressed at an acclimation temperature of 10 and 20 degrees C, but not at 30 degrees C. In contrast, MYHS30 mRNA was strongly expressed at all acclimation temperatures. The predominant MYH transcripts found in adult slow muscle and in embryos at hatching were expressed in adult fast muscle at some acclimation temperatures but not others. A MYH DNA clone was isolated from the cardiac muscle of 10 degrees C-acclimated adult fish (MYHcard). MYHcard mRNA was first detected at 61 h.p.f., but strong signals were only observed in the adult myocardium. The present study has therefore revealed a complex pattern of expression of MYH genes in relation to developmental stage, muscle type and acclimation temperature. None of the skeletal muscle MYHs identified so far was strongly expressed during the late juvenile stage, indicating further developmentally regulated members of the MYH II gene family remain to be discovered.

[Roles of reader's feelings in understanding narratives: forefeel, empathy, and a sense of strangeness].

This study examined the cross-domain, anticipatory, and self-referential roles of affect (Miall, 1989) in reading short mystery novels. In Experiment 1, undergraduate students either read an entire story once or read the same story twice. During the first reading, these participants read only the first part of the story, and during the second reading, they read the entire story. They rated the importance of each sentence. Then they either described a prediction or their impression of the story. Finally, they created a title. In Experiment 2, the first group rated the importance of each sentence. The second group rated the feelings of understanding: forefeel, empathy, and a sense of strangeness. The results of both experiments showed that the relative importance of sentences shifted between first reading and second reading. As they read the end of the story, empathy increased but a sense of strangeness decreased. The results of description showed that the readers updated their interpretations in order to understand the story correctly.

Temperature and the expression of myogenic regulatory factors (MRFs) and myosin heavy chain isoforms during embryogenesis in the common carp Cyprinus carpio L.

Embryos of the common carp, Cyprinus carpio L., were reared from fertilization of the eggs to inflation of the swim bladder in the larval stage at 18 and 25 degrees C. cRNA probes were used to detect transcripts of the myogenic regulatory factors MyoD, Myf-5 and myogenin, and five myosin heavy chain (MyHC) isoforms during development. The genes encoding Myf-5 and MyoD were switched on first in the unsegmented mesoderm, followed by myogenin as the somites developed. Myf-5 and MyoD transcripts were initially limited to the adaxial cells, but Myf-5 expression spread laterally into the presomitic mesoderm before somite formation. Two distinct bands of staining could be seen corresponding to the cellular fields of the forming somites, but as each furrow delineated, Myf-5 mRNA levels declined. Upon somite formation, MyoD expression spread laterally to encompass the full somite width. Expression of the myogenin gene was also switched on during somite formation, and expression of both transcripts persisted until the somites became chevron-shaped. Expression of MyoD was then downregulated shortly before myogenin. The expression patterns of the carp myogenic regulatory factor (MRF) genes most-closely resembled that seen in the zebrafish rather than the rainbow trout (where expression of MyoD remains restricted to the adaxial domain of the somite for a prolonged period) or the herring (where expression of MyoD persists longer than that of myogenin). Expression of two embryonic forms of MyHC began simultaneously at the 25-30 somite stage and continued until approximately two weeks post-hatch. However, the three adult isoforms of fast muscle MyHC were not detected in any stage examined, emphasizing a developmental gap that must be filled by other, as yet uncharacterised, MyHC isoform(s). No differences in the timing of expression of any mRNA transcripts were seen between temperature groups. A phylogenetic analysis of the MRFs was conducted using all available full-length amino acid sequences. A neighbour-joining tree indicated that all four members evolved from a common ancestral gene, which first duplicated into two lineages, each of which underwent a further duplication to produce Myf-5 and MyoD, and myogenin and MRF4. Parologous copies of MyoD from trout and Xenopus clustered closely together within clades, indicating recent duplications. By contrast, MyoD paralogues from gilthead seabream were more divergent, indicating a more-ancient duplication.

Empathy, spatial and verbal abilities characterize one who can best describe a route.

We investigated the characteristics of people who provided the most intelligible description of a route. 18 participants were asked to describe the route on their own campus. Then, participants completed the Japanese Wechsler Adults Intelligence Scale-Revised and the Interpersonal Reactivity Index. Subsequently, 72 raters 36 university students who were familiar with the environment and 36 students who were unfamiliar) were asked to read and rate each of the 18 descriptions on a 7-point scale based on intelligibility of the verbal descriptions. Analysis indicated that a person who can provide the most intelligible description of the route has a higher empathic concern, which is necessary to consider intelligibility of the description to the addressees as well as higher spatial abilities, which are relevant to spatial representation of the route.