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1.
J Oral Rehabil ; 2024 Jul 23.
Article in English | MEDLINE | ID: mdl-39044315

ABSTRACT

BACKGROUND: Oral and social problems can exacerbate long-term care. Understanding the relationship between social aspects and oral hypofunction can help identify high-risk factors for long-term care. OBJECTIVE: This study aimed to investigate the social aspects of oral hypofunction among medical outpatients. METHODS: This retrospective cross-sectional study included patients who visited an outpatient clinic for frailty. The oral function was assessed using seven items: oral hygiene, occlusal force, masticatory function, tongue-lip motor function, tongue pressure, oral dryness and swallowing function. Participants with three or more functional declines were classified as having 'oral hypofunction'. Social aspects were assessed using 21 items in four categories: general resources, basic social needs, social resources and social behaviours/activities. We analysed the relationship between oral hypofunction and each social aspect. RESULTS: A total of 316 participants (age 78.5 ± 6.4 years) were included, and 128 (41%) had oral hypofunction. Participants with oral hypofunction were significantly more likely to have limited education, require long-term care, not use transportation, depend on others for shopping, not participate in events, lack association membership and not engage in charity or volunteer work. After logistic regression analysis, 'long-term care' and 'no association membership' remained significantly associated with oral hypofunction (odds ratios 2.3, 2.3; 95% CI 1.1-5.0, 1.1-4.7, respectively). CONCLUSION: Participants with oral hypofunction faced challenges in 'general resources' and 'social behaviour/activities', which were linked to oral function issues. Future longitudinal studies are necessary to further investigate this relationship.

2.
Nihon Ronen Igakkai Zasshi ; 61(2): 155-162, 2024.
Article in Japanese | MEDLINE | ID: mdl-38839314

ABSTRACT

AIM: The coronavirus disease (COVID-19) pandemic has caused significant disruptions in various aspects of daily life. The Japanese Government declared a state of emergency in April 2020, which resulted in reduced physical activity. This study investigated the impact of these lifestyle changes by generation among outpatients with cardiovascular disease (CVD). METHODS: In autumn 2020, we conducted a questionnaire survey of 1,156 CVD outpatients who visited the Department of Cardiology at our institution. The survey collected data on physical activities and changes in daily behaviors over the course of the COVID-19 pandemic. Participants were classified into 3 age groups: middle-aged (n=114, ≤64 years old), semi-old (n=330, aged 65-74 years old), and old (n=712, ≥75 years old). The number of steps per day and sedentary time per day were compared between autumn 2019 and 2020, over the course of the pandemic. RESULTS: In autumn 2020, the number of steps per day was significantly decreased and sedentary time significantly increased in all age groups compared to the pre-pandemic levels. However, there were no significant differences in the extent of changes in steps per day or sedentary time over the study period across all age groups. Regarding changes in daily behaviors, only the old-age group reported a decline in volunteering and reduced utilization of daycare services. CONCLUSIONS: The COVID-19 pandemic has resulted in changes in daily activities and lifestyles across all age groups. Because lifestyle patterns differ across generations, it may be necessary to implement age-specific interventions and procedures.


Subject(s)
COVID-19 , Cardiovascular Diseases , Life Style , Outpatients , Pandemics , Humans , COVID-19/epidemiology , Aged , Cardiovascular Diseases/epidemiology , Middle Aged , Female , Male , Surveys and Questionnaires , Aged, 80 and over , Exercise , Adult , Japan/epidemiology
3.
J Cardiovasc Dev Dis ; 11(5)2024 Apr 25.
Article in English | MEDLINE | ID: mdl-38786955

ABSTRACT

BACKGROUND: Recent studies have investigated the effects of exercise on the functional capacity of older adults; training with a balance exercise assist robot (BEAR) effectively improves posture. This study compared the clinical safety and efficacy of training using BEAR video games to conventional resistance training in older adults with cardiovascular disease (CVD). METHODS: Ninety patients (mean age: 78 years) hospitalized due to worsening CVD were randomized to cardiac rehabilitation (CR) Group R (conventional resistance training) or Group B (training using BEAR). After appropriate therapy, patients underwent laboratory testing and functional evaluation using the timed up-and-go test (TUG), short physical performance battery (SPPB), and functional independence measure (FIM) just before discharge and 4 months after CR. The rates of CVD readmission, cardiac death, and fall-related fractures were monitored. RESULTS: BEAR had no adverse effects during exercise. At 4 months, TUG and SPPB improved significantly in both groups, with no significant difference between them. FIM motor and the Geriatric Nutritional Risk Index were significantly improved in Group B versus Group R. There was no significant difference in cardiac events and fall-related fractures between the two groups. CONCLUSION: CR with BEAR is safe and comparable to conventional resistance training for improving balance in older adults with CVD.

5.
Nutrition ; 124: 112438, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38657417

ABSTRACT

OBJECTIVES: In this study, we aimed to investigate the prevalence of oral frailty and explore its relationship with oral function and sarcopenia among older outpatients. MATERIALS AND METHODS: In this cross-sectional study, we retrospectively included older patients who visited a frailty outpatient clinic. We assessed total oral frailty employing, among other measures, oral diadochokinesis (/ta/ sound) for tongue-lip movement and tongue pressure. Patients who did not meet the cut-off values for three or more of these were classified as having oral frailty. Sarcopenia was assessed according to Asian Working Group for Sarcopenia 2019 criteria and analyzed for the relationship with oral function. RESULTS: The mean ± standard deviation age of the 111 patients was 77.2 ± 5.7 y; 63 were women (57%). Fifteen patients (14%) had either sarcopenia or dynapenia. The overall prevalence of oral frailty was 38%, with no significant difference in its prevalence between the sarcopenia/dynapenia group (44%) and the robust (no sarcopenia/dynapenia) group (35%). The following oral function assessments significantly differed between the sarcopenia/dynapenia group and the robust group: median (interquartile range) total oral frailty score, 2 (2-4) and 2 (1-3) (P = 0.019); tongue-lip motor function, 5.4 ± 1.2 and 5.9 ± 1.2 times/s (P = 0.049); and tongue pressure, 27.3 ± 8.5 kPa and 31.7 ± 8.0 kPa (P = 0.009). CONCLUSIONS: Approximately 40% of patients exhibited a decline in oral function regardless of the presence of sarcopenia. Sarcopenia and dynapenia may particularly affect tongue function. Although assessing patients for sarcopenia is crucial, separate evaluations of oral function should also be considered.


Subject(s)
Frail Elderly , Frailty , Geriatric Assessment , Outpatients , Sarcopenia , Tongue , Humans , Sarcopenia/epidemiology , Sarcopenia/complications , Cross-Sectional Studies , Female , Male , Aged , Frailty/epidemiology , Frailty/complications , Retrospective Studies , Prevalence , Outpatients/statistics & numerical data , Aged, 80 and over , Tongue/physiopathology , Geriatric Assessment/methods , Geriatric Assessment/statistics & numerical data , Frail Elderly/statistics & numerical data , Oral Health/statistics & numerical data
6.
Eur Geriatr Med ; 2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38438830

ABSTRACT

PURPOSE: This study aimed to investigate how intrinsic capacity (IC) deficit is associated with oral functional decline. METHODS: This cross-sectional study enrolled older adults at a research hospital frailty clinic between July 2021 and May 2023. IC evaluation included the locomotion, cognition, vitality, psychology, and sensory domains. Criteria for deficits were established within each domain, and the number of IC deficit domains was calculated for each patient. Oral function assessment included oral hygiene, oral dryness, occlusal force, tongue-lip motor function, tongue pressure, masticatory function, and swallowing function. Patients who met three or more criteria were classified into the oral hypofunction (OHF) group. Univariate and multivariate logistic regression analyses were performed to investigate the relationship between IC deficit and OHF. RESULTS: Of 222 included patients (mean age 78.3 ± 6.3 years; 39.6% men), 105 (47.3%) met the criteria for OHF. This OHF group showed a significantly higher prevalence of locomotion, cognition, psychology, and sensory domain deficits than the normal oral function group. Multivariate analysis adjusted for age and sex revealed a significant association between IC deficits and OHF (odds ratio [OR], 1.33; 95% confidence interval [CI] 1.04-1.70). A significant association was also observed between the locomotion domain and OHF (OR, 2.06; 95% CI 1.13-3.76). CONCLUSION: This study highlights the potential relationship between the number of IC domain deficits and oral functional decline, with the most significant domain being locomotion. Furthermore, it suggests a possible link between sensory and oral function.

7.
Geriatr Gerontol Int ; 24 Suppl 1: 102-109, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37746748

ABSTRACT

AIM: This study aimed to investigate the assessment tools dementia specialists use in clinical practice, reasons for their use and assessment-related factors. METHODS: A questionnaire survey was carried out from 15 September 2021 to 20 October 2021 among 1858 dementia specialists in Japan, with responses obtained via mail or using a Web form accessed via a Web address. RESULTS: Of the 1858 specialists who were sent the questionnaire, 574 responded, yielding a response rate of 32.2%. Almost all respondents stated that the main purposes of neuropsychological testing were to identify the pathophysiology and aid diagnosis. Most respondents identified behavioral and psychological symptoms of dementia as important factors for assessment. The most commonly used tests were the Hasegawa Dementia Scale-Revised and Mini-Mental State Examination, often used as screening tools. The Mini-Mental State Examination, Clock Drawing Test and Cube Copying Test were common assessments carried out directly by specialists. Quality of life and burden of care were less commonly assessed. CONCLUSIONS: Despite the main purpose of carrying out neuropsychological tests on dementia patients is to "understand the pathophysiology" and "aid in diagnosis," many assessment methods were chosen as screening methods carried out in a short time during clinic hours. The lack of evaluation of care burden and QOL, considered important by specialists, is an issue for the future in treating people with dementia, a life disability. Geriatr Gerontol Int 2024; 24: 102-109.


Subject(s)
Dementia , Humans , Dementia/diagnosis , Dementia/psychology , Quality of Life , Japan , Neuropsychological Tests , Mental Status and Dementia Tests
8.
Arch Gerontol Geriatr ; 118: 105305, 2024 03.
Article in English | MEDLINE | ID: mdl-38056104

ABSTRACT

OBJECTIVES: This study examined the diagnostic concordance and characteristics of the oral frailty five-item checklist (OF-5) and oral hypofunction (OHF). MATERIALS AND METHODS: In this extensive cross-sectional study, older adults were enrolled at a research hospital frailty clinic between July 2021 and July 2023. The diagnosis of OF-5 and OHF was conducted using the proposed assessment methods. The concordance rate of these criteria was determined by evaluating each patient and calculating the number of patients meeting each diagnosis's criteria. Patients who tested negative for the OF-5 and OHF criteria and those who met only one criterion were included in the OF-5 and OHF characteristic analysis. RESULTS: A total of 248 patients were included in the analysis (mean age 77.6 ± 6.8 years; 36.7 % men). Among the patients, 114 (46.0 %) and 116 (46.8 %) met the OF-5 and OHF criteria, respectively. Furthermore, 78 (31.5 %) participants met both the OF-5 and OHF criteria, whereas 96 (38.7 %) did not meet either criterion. Moreover, 36 (14.5 %) and 38 (15.3 %) patients tested exclusively positive for OF-5 and OHF, respectively. The OF-5-positive and OHF-negative group showed an older age, lower Mini Nutritional Assessment Short Form score, and higher fall risk than the OF-5-negative and OHF-negative group. However, no significant differences were observed between the OF-5-negative and OHF-positive and OF-5-negative and OHF-negative groups. CONCLUSION: The concordance rate of OF-5 and OHF diagnoses was 70.2 %. OF-5 showed a potential association with nutritional status and fall risk. Further longitudinal analyses are warranted to explore our study results.


Subject(s)
Frailty , Male , Humans , Aged , Aged, 80 and over , Female , Checklist , Cross-Sectional Studies , Nutritional Status , Geriatric Assessment/methods , Frail Elderly
9.
Nutrients ; 15(16)2023 Aug 12.
Article in English | MEDLINE | ID: mdl-37630750

ABSTRACT

Dysphagia is a syndrome of abnormal eating function resulting from a variety of causative diseases, and is associated with malnutrition. To date, the swallowing function has been difficult to examine without the use of invasive and expensive methods, such as the videofluorographic swallowing study or fiberoptic endoscopic evaluation of swallowing. In recent years, progress has been made in the clinical application of ultrasound equipment for the evaluation of body compositions near the body surface, including the assessment of nutritional status. Ultrasound examination is a noninvasive procedure and relatively inexpensive, and the equipment required is highly portable thanks to innovations such as wireless probes and tablet monitoring devices. The process of using ultrasound to visualize the geniohyoid muscle, digastric muscle, mylohyoid muscle, hyoid bone, tongue, masseter muscle, genioglossus muscle, orbicularis oris muscle, temporalis muscle, pharynx, esophagus, and larynx, and the methods used for evaluating these structures, are provided in this study in detail. This study also aims to propose a protocol for the assessment of swallowing-related muscles that can be applied in real-world clinical practice for the diagnosis of sarcopenic dysphagia, which can occur in elderly patients with sarcopenia, and has received much attention in recent years.


Subject(s)
Deglutition Disorders , Fractures, Bone , Aged , Humans , Deglutition , Deglutition Disorders/diagnostic imaging , Body Composition , Facial Muscles
10.
BMC Geriatr ; 23(1): 433, 2023 07 13.
Article in English | MEDLINE | ID: mdl-37442988

ABSTRACT

BACKGROUND: The spread of the novel severe acute respiratory syndrome coronavirus 2 infection has been prolonged, with the highly contagious Omicron variant becoming the predominant variant by 2022. Many patients admitted to dedicated coronavirus disease 2019 (COVID-19) wards (COVID-19 treatment units) develop disuse syndrome while being treated in the hospital, and their ability to perform activities of daily living declines, making it difficult for hospitals to discharge them. This study aimed to investigate the relationship between the degree of frailty and home discharge of patients admitted to a COVID-19 treatment units. METHODS: This study retrospectively examined the in-patient medical records of 138 patients (82.7 ± 7.6 years old) admitted to a COVID-19 treatment unit from January to December 2022. The end-point was to determine the patients' ability to be discharged from the unit directly to home; such patients were classified into the 'Home discharge' group and compared with those in the 'Difficulty in discharge' group. The degree of frailty was determined based on the Clinical Frailty Scale (CFS), and the relationship with the endpoint was analysed. A receiver operating characteristic (ROC) curve was created and the cut-off value was calculated with the possibility of home discharge as the state variable and CFS as the test variable. Logistic regression analysis was conducted with the possibility of home discharge as the dependent variable and CFS as the independent variable. RESULTS: There were 75 patients in the Home discharge group and 63 in the Difficulty in discharge group. ROC analysis showed a CFS cut-off value of 6 or more, with a sensitivity of 70.7% and a specificity of 84.1%. The results of the logistic regression analysis showed a significant correlation between possibility of home discharge and CFS even after adjusting for covariates, with an odds ratio of 13.44. CONCLUSIONS: Based on the evaluation of the degree of frailty conducted in the COVID-19 treatment unit, it was possible to accurately predict whether a patient could be discharged directly to home after treatment CFS could be an effective screening tool to easily detect patients requiring ongoing hospitalisation even after the acute phase of treatment.


Subject(s)
COVID-19 , Frailty , Humans , Aged , Aged, 80 and over , Frailty/diagnosis , Frailty/epidemiology , COVID-19/epidemiology , Retrospective Studies , SARS-CoV-2 , Activities of Daily Living , COVID-19 Drug Treatment , Frail Elderly , Hospitalization
13.
J Phys Ther Sci ; 34(11): 715-719, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36337216

ABSTRACT

[Purpose] Various types of Gait Exercise Assist Robot (GEAR) have been developed recently, some of which have enabled early improvement in patients with stroke. However, none has yet resulted in independent walking in these patients. Hence, we conducted an exploratory study of the effect of GEAR on achieving independent walking in stroke patients. [Participants and Methods] The participants were 16 patients with severe stroke. We evaluated patients' ability to walk independently after GEAR training. The outcome measure was Stroke Impairment Assessment Set (SIAS) motor score (Hip Flexion, Knee Extension, Foot Pat, Abdominal and Verticality). Differences in five SIAS motor scores were compared between the independent and non-independent walking groups. [Results] There was statistically significant difference between the groups in terms of Verticality among the 5 SIAS items used in the present research . Verticality of SIAS score of 1 was the cut-off value for distinguishing walking independence. [Conclusion] Verticality of SIAS may be a marker of potential walking independence that can be used in rehabilitation plans using walking-assist robots in patients with stroke.

15.
J Cardiovasc Dev Dis ; 9(6)2022 Jun 12.
Article in English | MEDLINE | ID: mdl-35735820

ABSTRACT

We examined whether adding robot-supported balance exercises to cardiac rehabilitation improves the ability to balance in older adults with cardiovascular disease (CVD). We conducted a prospective study in 52 older adults who had been hospitalized for worsening CVD. Once weekly for four months, for a total of sixteen sessions as outpatients, the subjects used a Balance Exercise Assist Robot (BEAR) to perform balance exercises and an ergometer for aerobic exercises. Participants' mean age was 76.9 ± 6.8 years (range, 65−95 years), and their mean brain natriuretic protein level was 164.0 ± 190.0 pg/mL. After the intervention, participants showed significant improvements in gait speed (before, 1.06 ± 0.33 m/s; after, 1.23 ± 0.30 m/s; p < 0.001), Short Physical Performance Battery score (before, 10.02 ± 2.25; after, 10.88 ± 1.79; p ˂ 0.001), timed up-and-go (before, 11.11 ± 5.07 s; after, 9.45 ± 3.45 s; p ˂ 0.001), and knee extension (before, 26.97 ± 11.78 kgf; after, 30.13 ± 13.04 kgf; p = 0.001). Cardiac rehabilitation including exercises using BEAR improved physical functioning and the ability to balance in older adults with CVD. Frail and prefrail patients improved, whereas robust ones did not change.

16.
Geriatr Gerontol Int ; 21(10): 919-925, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34382724

ABSTRACT

AIM: As the number of frail and disabled older people increases in Japan, systems are needed to detect efficiently the populations at high risk in need of care and to allow early intervention. We investigated cut-off values for those physical functioning assessments adopted by AWGS 2019 associated with new or worsening long-term care insurance (LTCI) certification. METHODS: We recruited 497 outpatients from our locomotive syndrome-frailty clinic. After excluding patients who had undergone orthopedic surgery right after assessment or ≤65 years old, 233 patients (mean age, 78 ± 6 years) were included. Logistic regression analysis was performed using new certification for LTCI services need or deterioration of care status in 1 year as the dependent variable and physical function and other assessments as independent variables. Next, we constructed receiver operating characteristic curves and calculated areas under the curve and optimal cut-off values. RESULTS: During 1-year follow-up, 37 patients (16%) obtained new certification for LTCI services need or deterioration of status. After adjusting, usual walking speed and Short Physical Performance Battery score were significantly associated with outcomes. With receiver operating characteristic curves, usual walking speed as the test variable showed an area under the curve of 0.740 with a cut-off of 0.92 m/s, whereas the Short Physical Performance Battery score showed an area under the curve of 0.737 with a cut-off score of 9. CONCLUSIONS: Slower walking speed and lower Short Physical Performance Battery score may predict new or worsening LTCI for older people. Geriatr Gerontol Int 2021; 21: 919-925.


Subject(s)
Frailty , Aged , Aged, 80 and over , Frail Elderly , Geriatric Assessment , Humans , Long-Term Care , Outpatients , Physical Functional Performance , Walking Speed
17.
Geriatr Gerontol Int ; 21(10): 900-906, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34363304

ABSTRACT

AIM: Decreased use of life spaces, as reflected in decreased Life-Space Assessment (LSA) scores, is associated with poor prognosis in older adults. The purpose of this study was to examine factors affecting the extent of life-space activities in older adults with cardiovascular disease. METHODS: We carried out a prospective observational study in 98 older adults (minimum age 65 years; mean age 79.5 ± 7.4 years) who were admitted to our hospital due to cardiovascular disease. Once their medical condition was stable, they underwent cardiopulmonary exercise testing, echocardiography and physical evaluation, and completed questionnaires. RESULTS: The LSA score was significantly associated with the ability to drive a car (driving 95.1 ± 21.1 points, not driving 60.4 ± 30.3 points, P < 0.001). In addition, LSA was significantly correlated with age; peak VO2 ; brain natriuretic peptide; and Short Physical Performance Battery, Geriatric Depression Scale and Mini-Mental State Examination scores. In a multiple regression analysis, Short Physical Performance Battery and driving a car were significantly associated with LSA (ß = 0.28, ß = 0.37, respectively). CONCLUSION: Assessment of motor function and social factors in addition to clinical cardiac function might be important to understand the complete context of life-space activity in older adults with cardiovascular disease. Geriatr Gerontol Int 2021; 21: 900-906.


Subject(s)
Automobile Driving , Cardiovascular Diseases , Activities of Daily Living , Aged , Aged, 80 and over , Automobiles , Cardiovascular Diseases/diagnosis , Geriatric Assessment , Humans , Natriuretic Peptide, Brain , Physical Functional Performance
18.
Analyst ; 144(13): 4045-4050, 2019 Jul 07.
Article in English | MEDLINE | ID: mdl-31157334

ABSTRACT

Two-photon excited fluorescence (TPEF) of rhodamine 6G (Rh6G) and tris(dibenzoylmethane) mono(5-aminophenanthroline) europium (Eu-TDPA) was measured using a pulsed diode laser head (<45 mW, 975 nm, 90 ps pulse width, 40 MHz). Fluorophores were cast on a glass slide modified with triangular silver nanoprisms. A photon-counting photomultiplier detected the TPEF of Rh6G on a glass substrate (1361 Hz) and on the nanoprisms (6322 Hz). On the other hand, Eu-TDPA did not exhibit TPEF on a glass substrate. TPEF was only observed when the extinction of the nanoprisms on the substrates was larger than 0.1. The nanoprisms enhanced the TPEF of these two fluorophores up to the detectable level using a low-power laser diode.

19.
Nihon Ronen Igakkai Zasshi ; 55(1): 65-73, 2018.
Article in Japanese | MEDLINE | ID: mdl-29503370

ABSTRACT

PURPOSE: We examined the factors related to life space and changes in the care level after one year in daycare center users. METHODS: The participants were 83 older adults (age, > 65 years; mean age, 79.5±6.8 years) with MMSE scores of ≥20, who could walk independently, who needed support (1-2) or care (1), and who underwent rehabilitation at a daycare center. The life space was evaluated by the Life Space Assessment (LSA). The subjects' basic information (i.e., age, medical history.) was collected, and their physical function (i.e., grip strength, timed up and go test [TUG]), mental function (i.e., vitality, fear of falls), and social function (i.e., friends, hobbies, public transportation) were assessed to investigate the factors associated with their LSA scores. In addition, a follow-up survey was conducted on the care level at approximately one year later. RESULTS: A multiple regression analysis indicated that TUG scores (ß=-0.33), hobbies (ß=0.30), friends (ß=0.29), public transportation (ß=0.26), and grip strength (ß=0.24) were related to the life space. Next, the participants were divided into LSA-high and LSA-low groups, and changes in the care level (improvement, maintenance, deterioration) at approximately one year after the initial assessment were examined using a chi-squared test. A significant difference was observed in the distribution of the groups (p=0.03). CONCLUSIONS: Multiple factors were related to the life space. Moreover, it is possible that improvements in the level of care may be achieved by improving the life space.


Subject(s)
Activities of Daily Living , Aged , Aged, 80 and over , Female , Geriatric Assessment , Humans , Male , Social Behavior
20.
Plant Signal Behav ; 8(1): e22510, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23123450

ABSTRACT

Autophagy has been shown to play essential roles in the growth, development and survival of eukaryotic cells. However, simple methods for quantification and visualization of autophagic flux remain to be developed in living plant cells. Here, we analyzed the autophagic flux in transgenic tobacco BY-2 cell lines expressing fluorescence-tagged NtATG8a as a marker for autophagosome formation. Under sucrose-starved conditions, the number of punctate signals of YFP-NtATG8a increased, and the fluorescence intensity of the cytoplasm and nucleoplasm decreased. Conversely, these changes were not observed in BY-2 cells expressing a C-terminal glycine deletion mutant of the NtATG8a protein (NtATG8aΔG). To monitor the autophagic flux more easily, we generated a transgenic BY-2 cell line expressing NtATG8a fused to a pH-sensitive fluorescent tag, a tandem fusion of the acid-insensitive RFP and the acid-sensitive YFP. In sucrose-rich conditions, both fluorescent signals were detected in the cytoplasm and only weakly in the vacuole. In contrast, under sucrose-starved conditions, the fluorescence intensity of the cytoplasm decreased, and the RFP signal clearly increased in the vacuole, corresponding to the fusion of the autophagosome to the vacuole and translocation of ATG8 from the cytoplasm to the vacuole. Moreover, we introduce a novel simple easy way to monitor the autophagic flux non-invasively by only measuring the ratio of fluorescence of RFP and YFP in the cell suspension using a fluorescent image analyzer without microscopy. The present in vivo quantitative monitoring system for the autophagic flux offers a powerful tool for determining the physiological functions and molecular mechanisms of plant autophagy induced by environmental stimuli.


Subject(s)
Autophagy , Cytoplasm , Nicotiana/physiology , Phagosomes , Plant Cells/physiology , Plant Proteins/metabolism , Vacuoles , Cell Line , Fluorescence , Plants, Genetically Modified , Recombinant Fusion Proteins
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