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Introduction: Due to the ongoing outbreak of the coronavirus disease 2019 (COVID-19), it is currently difficult to conduct in-person exercise classes. We thus started the program of an online physical exercise with musical accompaniment. Several interesting differences were found in the characteristics of the online participants compared with our previous in-person interventions. Participants and Methods: The total number of subjects was 88 (71.2 ± 4.9 years old; male 42, female 46). The questionnaire included the attributes of the participants, the perceived advantages of the exercise classes, and the presence or absence of noticeable changes in cognitive and physical function after participating in the classes. Results: The personal computers used to attend the online classes were operated by the participants themselves. About 42% of the participants felt that their sense of day of the week and volition were improved by attending the exercise classes for 3 months. The most frequent answer to the reason for participation was because it was free (81.8%). The second most frequent answer was because the classes were held online (75.0%). Almost half of the participants answered that they would not participate if it was held in person because of the risk of COVID-19 infection (75.0%) and the difficulty getting to the site where the exercise classes were held (59.1%). Conclusion: Online physical exercise with musical accompaniment improved the perceived orientation, volition, activity, exercise habits, and health condition in 30-40% of the participants and also stimulated greater participation by males compared to classes held in person.

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Objective: The spread of coronavirus disease (COVID-19) has limited the implementation of face-to-face non-pharmacological treatment for the prevention of dementia. As a result, online non-pharmacological treatment has become increasingly important. In this study, we used an online conferencing system to implement an online version of a physical exercise program with music, and examined its effect on cognitive function. Methods: The participants were 114 healthy older adults [63 men and 51 women; mean age of 70.7 years (standard deviation = 4.6)]. Seventy-five participants were allocated to the physical exercise with music group (60 min, once a week, total 20 sessions), while the remaining 39 participants were assigned to the control group, and only underwent the examinations. In the physical exercise with music group, we performed neuropsychological examinations and brain tests both before and after the exercise program. Neuropsychological tests included the Mini-Mental State Examination, Raven's Colored Progressive Matrices (RCPM), the Rivermead Behavioral Memory Test, graphic imitation, word fluency (WF) (animal names and initial sounds), and the Trail Making Test-A/B. As an assessment of brain function, we developed an online examination of subtle cognitive decline, including tests of number and word memory, spatial grasp, the N-back task, and change inference. Results: In the N-back task, the physical exercise with music group improved significantly relative to the control group (p = 0.008). Discussion: The present findings suggest that the online version of the physical exercise with music program improved working memory, which mainly involves the frontal lobe.

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INTRODUCTION: We have developed an online cognitive test (Brain Assessment [BA]) which exhibits moderate correlations with established neuropsychological tests. We made a short version of the BA that requires only 10 min and investigated correlations with the original version. PARTICIPANTS AND METHODS: Three hundred and twenty-three subjects (26-82 years old, mean age 41.9 years) were recruited via the internet and performed the original version of the BA online, and, for the analyses of the original and short version, the results of the whole and of the first 2 min were utilized, respectively. Correlations were evaluated between both versions, and 144 subjects agreed to be interviewed about BA utilization. RESULTS: Correlations between the short and original versions of the BA were strong (memory of numbers, r = 0.72: total, r = 0.73) or moderate (memory of words, r = 0.66; visuospatial, r = 0.42; working memory, r = 0.53; judgment, r = 0.54). We found a strong negative correlation between the "memory of words" subtest and age (short version, r = -0.85; original version, r = -0.88). Regarding the interview, most wanted to be assessed by the short version every year. In the case of a concerning score, they would then want to be examined by the original version of the BA and/or more specific neuropsychological tests. DISCUSSION: We found that the short and the original versions of the BA exhibited strong correlations, and the correlation coefficients between age and subtest scores were almost the same between the two versions. CONCLUSION: The short version of the BA will be useful to perform cognitive assessments in the clinic.

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INTRODUCTION: There are several problems with standard in-person neuropsychological assessments, such as habituation, necessity of human resources, and difficulty of in-person assessment under societal conditions during the outbreak of coronavirus disease 2019. Thus, we developed an online cognitive test (the Brain Assessment [BA]). In this study, we investigated the correlation between the results of the BA and those of established neuropsychological tests. PARTICIPANTS AND METHODS: Seventy-seven elderly persons (mean 71.3 ± 5.1 years old; range 65-86; male:female = 45:32) were recruited through the internet. Correlations were evaluated between the BA and the following widely used neuropsychological tests: the mini-mental state examination (MMSE), the Raven's colored progressive matrices (RCPM), the logical memory I and II of the Rivermead Behavioral Memory Test, the word fluency (WF) test, and the Trail-Making TestA/B. RESULTS: We found moderate correlations between the total cognitive score of the BA and the total score of the MMSE (r = 0.433, p < 0.001), as well as between the total BA score and the total RCPM score (r = 0.582, p < 0.001) and time to complete the RCPM (r = 0.455, p < 0.001). Moderate correlations were also observed between the cognitive score of the memory of words BA subtest and the LM-I (r = 0.518, p < 0.001), the mental rotation subtest and figure drawing (r = 0.404, p < 0.001), the logical reasoning subtest and total RCPM score (r = 0.491, p < 0.001), and the memory of numbers and words subtests and WF (memory of numbers and total WF: r = 0.456, p < 0.001; memory of words and total WF: r = 0.571, p < 0.001). DISCUSSION: We found that the BA showed moderate correlations between established neuropsychological tests for intellect, memory, visuospatial function, and frontal function. The MMSE and the RCPM reflect Spearman's s-factor and g-factor, respectively, and thus the BA also covered both factors. CONCLUSION: The BA is a useful tool for assessing the cognitive function of generally healthy elderly persons.

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INTRODUCTION: It is well-known that cognitive function declines with age. In order to detect changes in cognitive function, cognitive tests should be performed repeatedly. Currently existing cognitive tests come in only a single version, so the subject is likely to remember the contents with repeated testing. And, under the outbreak of coronavirus disease 2019 (COVID-19), in-person assessment should be avoided. This study was performed to develop a new cognitive test (brain assessment, BA) that has 5 versions and can be performed on a personal computer (PC) through the Internet. MATERIALS AND METHODS: Five thousand subjects performed the online BA, which consisted of 5 subtests: number memory, word memory, mental rotation test, N-back test, and judgment test. We standardized the raw scores (cognitive scores, CSs) using mean and standard deviation, which were 50 and 10, respectively. Then, we calculated the mean CS for each sex and age, plotted the relationships between ages and mean CSs on figures, and calculated the formula of cognitive changes during normal aging. RESULTS: The CSs of all subtests decreased with aging. The regression coefficient was from -0.31 to -0.45. It is noteworthy that in most subtests, the CSs started to increase at 85 years of age. DISCUSSION: Our BA has 5 versions and can be done on a PC using the Internet. We tested the BA in a large number of subjects, and the standard values of CSs were measured in individuals up to 89 years of age. By performing this test repeatedly, subjects can evaluate the degree of their cognitive decline. If the rate of cognitive decline is greater than that predicted using the normalized formula, the subjects can undertake strategies to improve their control of lifestyle-related diseases or other habits of daily living. CONCLUSION: The BA can be easily taken online using a PC, and its scores linearly declined with normal aging. The BA will be useful for detecting longitudinal cognitive changes and comparing them to the pattern seen in normal aging.

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A one-step, single tube, real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting sequences of the untranslated region in the genome of hepatitis A virus (HAV). The RT-LAMP assay reported in this study was very simple and rapid; the HAV-specific amplification was obtained in 50 min under isothermal conditions at 62.5 degrees C by employing a set of seven primers. The RNAs of three cell-adapted HAV strains belonging to different subgenotypes (IA, IB and IIIB) were equally well amplified. The detection limits of the RT-LAMP assay for these HAV strains were 0.4-0.8 focus forming units (FFU)/reaction. The results of the calibration using the WHO international standard indicated that the RT-LAMP assay had similar sensitivity to the conventional RT-PCR method. A comparison of the results from the RT-LAMP and the LightCycler PCR assay using clinical samples in feces revealed that the findings were similar between the two methods. Although several genotypes remain to be tested, it is concluded that the new real-time RT-LAMP assay is very suitable for detection and quantitation of most prevalent genotypes of HAV in diagnostic laboratories.

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Mumps virus is still circulating and annual mumps outbreaks occur with fluctuating magnitudes in Japan. Aseptic meningitis has been reported after vaccination and it would be of importance to determine whether this was related to the vaccination. The objective of this study was to develop a sensitive, specific and rapid diagnostic method for the differentiation of the Hoshino vaccine strain from circulating wild types. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method of the hemagglutinin neuraminidase (HN) region for the detection of mumps virus genome from clinical samples. The typical ladder pattern disappeared after the LAMP products of the Hoshino vaccine strain were digested with ScaI, but those of wild types were not cut by ScaI. We obtained 19 cerebro spinal fluids (CSF) from the patients with aseptic meningitis and 17 salivary swab samples from the patients with acute parotitis after mumps vaccination, in which one case was complicated with orchitis. Mumps virus genome was detected in 18 CSF samples and in all NPS by RT-LAMP. The Hoshino vaccine strain was identified in 16 out of 18 CSF RT-LAMP positives and in 11 out of 17 NPS samples and the remaining samples were identified as wild types. RT-LAMP followed by ScaI digestion is a sensitive, simple and rapid differential method and useful for laboratory surveillance for vaccine-adverse events.

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We developed a useful method for the detection of rubella virus genome RNA by reverse transcription loop-mediated isothermal amplification (RT-LAMP) and compared the sensitivity of RT-LAMP with that of other virological tests: reverse transcription-PCR (RT-PCR) and virus isolation. The rubella virus genome was amplified by RT-LAMP from clinical isolates obtained between 1987 and 2004 with similar sensitivities to the Takahashi vaccine strain. The detection limit of RT-LAMP was compared with that of RT-PCR using the Takahashi vaccine strain. We detected rubella virus genome material corresponding to 30 PFU/ml in a culture fluid sample by RT-LAMP within 60 min after the extraction of RNA with equal sensitivity to RT-nested PCR. The positive result rates of RT-LAMP, RT-PCR, and virus isolation were also compared using throat swabs obtained from patients who were clinically diagnosed with acute rubella virus infection in 2004 in Tochigi, Japan. Among nine patients with clinical rubella, the positive result rates were three/nine (33.3%) for virus isolation, six/nine (66.7%) for RT-PCR, and seven/nine (77.8%) for RT-LAMP. Consequently, RT-LAMP for rubella virus would be expected to be a reliable rapid diagnostic tool in the clinical setting.

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Annual seasonal outbreaks of respiratory syncytial virus (RSV) infection occur every winter. Most patients are diagnosed clinically by a rapid detection kit for RSV protein(s) from nasopharyngeal secretion (NPS), but some problems have been reported on the specificity and sensitivity of such rapid detection kits. To ratify these issues, a sensitive, specific, simple, and rapid molecular based diagnostic method is expected to be introduced and we have developed a method to detect the RSV genome of subgroups A and B independently by reverse transcription loop-mediated isothermal amplification (RT-LAMP). We detected the genomic RNA corresponding approximately to 0.1 TCID 50 in the sample by RT-LAMP for both RSV subgroups under isothermal condition within 60 min after extraction of RNA. Specific DNA amplification was monitored by a real-time turbidimeter and the quantity of RNA was calculated. The RSV genome was detected in 47 of 50 NPS by RT-LAMP, and in 42 by nested RT-PCR, whereas virus isolation was positive for 29 and enzyme-linked immunoassay (EIA) for 34. RSV subgroup A was detected in 25 by RSV RT-LAMP A, RSV subgroup B in 23 by RSV RT-LAMP B, and dual infection with RSV subgroups A and B was identified in one case. They were confirmed with digestion with a specific restriction enzyme, Bgl II. The results showed the potential clinical feasibility of RT-LAMP as a useful diagnostic tool for the detection of RSV with high sensitivity similar to nested RT-PCR.

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Approximately 20,000-30,000 measles patients were reported in a surveillance of infectious diseases because of low vaccine coverage of 80% in Japan. Among them, some were thought to be secondary vaccine failure (SVF) with generally mild or non-typical measles illness and sometimes became a source of further transmission. We have developed a new, sensitive, and rapid method to detect the measles virus genome by reverse transcription loop-mediated isothermal amplification (RT-LAMP). We examined 50 nasopharyngeal secretion (NPS) samples that were obtained during the 1999 outbreak and stored at -70 degrees C and fresh NPS, lymphocytes and sera from 11 patients in 2003. Total RNA was extracted from the samples and subjected to reverse transcription-polymerase chain reaction (RT-PCR) and RT-LAMP. We detected the genomic RNA corresponding to at least 0.01-0.04 TCID50, 30-100 copies in samples by RT-LAMP within 60 min after extraction of RNA, and all four genotypes isolated in Japan were equally amplified. Specific DNA amplification was monitored spectrophotometrically by real time turbidimeter and the quantity of RNA was calculated. Measles virus genome was detected in 44 of 50 stored NPS by RT-PCR and in 49 by RT-LAMP. The vaccine strain was discriminated from wild strains after sequencing the LAMP products. RT-LAMP is a useful rapid diagnostic method for the detection of measles virus without any special apparatus, showing higher sensitivity than RT-PCR, and expected to be applied for hospital-based infection control and for laboratory-based measles surveillance.

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Most mumps patients are clinically diagnosed without any virological examinations, but some diagnosed cases of mumps may be caused by other pathogens or secondary vaccine failure (SVF). To clarify these issues, a sensitive, specific, and rapid diagnostic method is required. We obtained 60 salivary swabs from 34 patients with natural infection during the course of the illness, 10 samples from patients with vaccine-associated parotitis, and 5 samples from patients with SVF. Total RNA was extracted and subjected to reverse transcription-PCR (RT-PCR) and loop-mediated isothermal amplification (LAMP) for genome amplification. We detected mumps virus RNA corresponding to 0.1 PFU by LAMP within 60 min after RNA extraction, with the same sensitivity as RT-nested PCR. Mumps virus was isolated in 30 of 33 samples within day 2, and mumps virus genome was amplified by LAMP in 32 of them. The quantity of virus titer was calculated by monitoring the time to reach the threshold of turbidity. The viral load decreased after day 3 and was lower in patients serologically diagnosed as having SVF with milder illness. Accuracy of LAMP for the detection of mumps virus genome was confirmed; furthermore, it is of benefit for calculating the viral load, which reflects disease pathogenesis.

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We studied gastric function in patients with functional dyspepsia, using capsules containing contrast medium. We were able to estimate intragastric capsule movement by this method. Here, we report gastric functions in patients with esophagitis. We were able to recognize gastric dysfunction (delayed gastric emptying) in these patients.

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