Single-Cell Transcriptome Identifies the Renal Cell Type Tropism of Human BK Polyomavirus.

BK polyomavirus (BKPyV) infection is the main factor affecting the prognosis of kidney transplant recipients, as no antiviral agent is yet available. A better understanding of the renal-cell-type tropism of BKPyV can serve to develop new treatment strategies. In this study, the single-cell transcriptomic analysis demonstrated that the ranking of BKPyV tropism for the kidney was proximal tubule cells (PT), collecting duct cells (CD), and glomerular endothelial cells (GEC) according to the signature of renal cell type and immune microenvironment. In normal kidneys, we found that BKPyV infection-related transcription factors P65 and CEBPB were PT-specific transcription factors, and PT showed higher glycolysis/gluconeogenesis activities than CD and GEC. Furthermore, in the BKPyV-infected kidneys, the percentage of late viral transcripts in PT was significantly higher than in CD and GEC. In addition, PT had the smallest cell-cell interactions with immune cells compared to CD and GEC in both normal and BKPyV-infected kidneys. Subsequently, we indirectly demonstrated the ranking of BKPyV tropism via the clinical observation of sequential biopsies. Together, our results provided in-depth insights into the renal cell-type tropism of BKPyV in vivo at single-cell resolution and proposed a novel antiviral target.

Difficulties and countermeasures in implementing age-friendly policies in primary health centers in Taiwan.

BACKGROUND: Taiwan is predicted to become a super-aged society by 2025, and primary health centers (PHCs) are set to play a crucial role in the care of older adults. The Taiwanese government has developed an age-friendly verification framework for PHC. The aims of this study were to explore the difficulties faced by PHC staff in the implementation of age-friendly policies and their solution strategies. METHODS: This study adopted a qualitative research method. The first stage involved conducting five focus groups with the responsible staff of PHCs (n = 41) that have been certified "age-friendly." The focus groups covered the effectiveness, difficulties, and resources of PHCs in regards to the introduction of age-friendly policies. In the second stage, in-depth interviews were conducted with executives of PHCs (n = 5), both certified and not certified as age-friendly, to further compare the difficulties faced by these two types of PHCs, thereby gaining perspectives for solution strategies. The principles of grounded theory were used for data analysis. RESULTS: Four major PHC strategies are employed in the promotion of age-friendliness. First, organizational management, through which managers apply management methods and analyze the present PHC-related health concerns; second, resource utilization, which refers to the tallying, linking, and integrating of resources; third, business operation process, in which work efficiency is improved through the combination of business operations and staff training; finally, hardware improvement, which is achieved through comprehensive cataloging of facility environments. CONCLUSION: The implementation of age-friendliness in PHCs requires the efforts of both the service units and government. With resources provided by the government, PHCs can integrate management methods, businesses operations, and essential resources. Moreover, PHC executives can lead their teams in promoting age-friendly policies, and closely monitor their effectiveness.

Analysis of drug sensitivity of Escherichia Coli O157H7.

Sensitive and rapid tests of Escherichia coli drug sensitivity is very important for health of human and animals. An E. coli immunosensor was built based on electrochemical detection and immune detection technologies, through pretreating screen-printed electrodes, and analyzing the optimal reaction concentration of antigen antibody binding with the AC impedance method. Based on the detection system combining the immunosensor and electrochemical workstation, tests were carried out to measure the accuracy of E. coli concentration and drug sensitivity, and error of the detection system was calibrated in accordance with data from the electrochemical workstation. E. coli O157:H7 can be detected in the range of 103 cfu/ml ~ 1012 cfu/ml, and the detection error controlled within 5%. Results from the electrochemical workstation and those from the detection device were consistent, and both demonstrated a greater inhibitory effect of antibiotics on E. coli than on Bacillus subtilis. The electrochemical detection system is highly efficient and accurate, and could be widely applied to E. coli drug sensitivity tests in clinical medicine.

Consideration factors of older adults seeking medical treatment at outpatient services in Taiwan.

BACKGROUND: Taiwan will become a super-aged society by 2025, leading to the more frequent use of outpatient services by older adults for medical treatment compared with other age groups. Understanding the outpatient service consideration factors of older adults seeking medical treatment can improve health care quality. This study explored the selection factors and crucial considerations of older adults for outpatient services. METHODS: Qualitative study was conducted. Purposive sampling was used to recruit 16 older adults over 65 years of age with chronic disease who were patients of an internal medicine department and regularly returned for checkups. Data including reasons for receiving medical treatment, factors affecting their choice of hospitals, and health care and environmental considerations were collected through structured interviews. RESULTS: The older adults identified four factors. (1) The care of doctors: The doctors possessed professional skill, allocate sufficient consultation time, and undertake effective communication. (2) The care of other medical professionals: Other medical professionals provided services in a cordial manner. (3) The accessibility and convenience of outpatient services: Convenient transportation and registration as well as short consultation wait time. (4) Environment and equipment: The hospital had the novel facilities and satisfactory barrier-free equipment. CONCLUSIONS: The older adults cared most about the adequacy of diagnosis and treatment by doctors and other medical professionals. In addition, they reported having higher satisfaction with hospitals that provide comprehensive medical facilities, fast and convenient medical procedures, and short wait times.

Working Experiences of Care Aides in Long-Term Care Institutions Following the Relaxation of COVID-19 Regulations in Taiwan: A Qualitative Study.

Care aides in long-term care (LTC) institutions care for older disabled residents at high risk for COVID-19. However, they experienced many stressors during the COVID-19 pandemic. This study aims to explore the working experiences of care aides in LTC institutions following the relaxation of COVID-19 regulations in Taiwan. This qualitative descriptive study included 20 care aides who had cared for residents with COVID-19. Data were obtained via semi-structured interviews. Caring for residents with COVID-19 and the difficulties, resources and teamwork, and impact of care aides' work on their lives were discussed. Consequently, four themes were identified. First, difficulties in care, which included physical limitations by protection, workload, and impact of work schedule on the lives of the care aides. Second, psychological impact, such as worry, social isolation, and burnout. Third, interpersonal relationships with supervisors, colleagues, residents, and their families. Fourth, infection control policy from the institution and government. When infection control policies were relaxed, care aides had difficulties in caring for residents; furthermore, their family and social lives were also affected. They were required to learn knowledge of and skills for COVID-19 management. Institutions were required to provide support in materials, care processes, environment, and management.

Differences in the healthcare needs of older adults attending primary health centers in urban and rural areas of Taiwan.

PURPOSE: As the older adult population in Taiwan continues to increase, primary health centers (PHCs) play a crucial role in geriatric care. This study explored the differences in the PHC experiences and usage needs of older adults in urban versus rural areas. METHODS: A qualitative study was conducted. Twenty-one older adults were recruited from PHCs in northern, central, southern, and eastern Taiwan. Semistructured interviews were used to collect data, and the interview guidelines included their reasons for visiting PHCs, the health-care services they received, their evaluation of the services, and the advantages and disadvantages of these centers. The data were analyzed using thematic content analysis. RESULTS: The PHC usage needs of older adults in urban areas differ from those of older adults in rural areas in the following 3 aspects: (1) Medical services: older adults in rural areas demand clinics specializing in various medical domains, mobile and home medical care, and case management, whereas those in urban areas demand mobile health examinations. (2) Environment and transportation: older adults in rural areas demand diverse medical equipment, shuttle services, and accessible facilities, whereas those in urban areas demand recreational facilities. (3) Active aging: older adults in rural areas demand health education courses, and those in urban areas demand diverse senior citizen courses as well as opportunities to volunteer and build interpersonal relationships. CONCLUSION: The older adults in urban and rural areas had different PHC usage needs. The older adults in rural areas generally focused on medical care and transportation services in PHCs, whereas those in urban areas generally focused on health promotion as a means of social participation and active aging.

Tenascin-C promotes the proliferation and fibrosis of mesangial cells in diabetic nephropathy through the ß-catenin pathway.

OBJECTIVE: To mechanistically assess the involvement of tenascin-C (TNC) in diabetic nephropathy (DN). METHODS: Renal specimens from DN patients were histopathologically examined, and their TNC expression patterns were evaluated via immunohistochemistry. Additionally, the hereditarily diabetic C57BL/KsJ db/db mice were induced to develop DN via adaptive feeding, and then their renal levels of TNC and ß-catenin were assessed via western blotting and immunohistochemistry. Furthermore, the TNC and ß-catenin levels in primary rat mesangial cells (RMCs) cultured with high glucose levels were assessed via western blotting. In parallel, RMCs cultured with TNC in the presence or absence of the ß-catenin blocker ICG-001 were analyzed for their fibronectin and collagen I levels via immunostaining, and for their fibronectin, α-SMA, vimentin, PDGFR-ß, PCNA, and ß-catenin levels via western blotting. RESULTS: The TNC levels in the specimens were associated with the pathological classification. In these DN specimens, TNC protein was highly detected in the MCs and slightly in the tubulointerstitium. Renal TNC (P < 0.05) and ß-catenin (P < 0.001) were upregulated in db/db vs. db/m mice. High-glucose treatment upregulated TNC (P < 0.01) and ß-catenin (P < 0.05) in RMCs. TNC treatment upregulated fibronectin (P < 0.05), α-SMA (P < 0.01), vimentin (P < 0.05), PCNA (P < 0.05), and ß-catenin (P < 0.05) in RMCs, as assessed via western blotting. Immunohistochemical analysis confirmed the fibronectin upregulation and showed collagen I upregulation. Western-blot results also showed that levels of fibronectin (P < 0.001), α-SMA (P < 0.01), vimentin (P < 0.001), PCNA (P < 0.05), PDGFR-ß (P < 0.05), and ß-catenin (P < 0.01) were lower in RMCs co-treated with TNC and ICG-001 than in TNC-treated cells. Immunofluorescence analysis confirmed the decreased fibronectin level and showed that the collagen I level was also decreased by ICG-001. CONCLUSION: TNC is upregulated in DN and induces MC proliferation and fibrosis through the ß-catenin pathway.

The Associations between Physical Activity, Functional Fitness, and Life Satisfaction among Community-Dwelling Older Adults.

Previous studies showed physical activity had benefits for older adults' life satisfaction, but the mechanism was unclear. This study aimed to investigate whether older adults with more physical activity had better functional fitness and life satisfaction, and whether functional fitness mediated the relationship between physical activity and life satisfaction. A cross-sectional study design was employed, and 623 older adults (73.71 ± 5.91 years) were recruited. Physical activity, functional fitness, life satisfaction, and demographic characteristics were collected. Compared with older adults with low physical activity, those with high (B = 0.41, p = 0.025) and moderate (B = 0.40, p = 0.041) physical activity had better life satisfaction; those with high physical activity had better lower limb muscle strength (B = 1.71, p = 0.001), upper (B = 2.91, p = 0.032) and lower (B = 3.12, p = 0.006) limb flexibility, cardiorespiratory endurance (B = 6.65, p = 0.008), and dynamic balance ability (B = −1.12, p < 0.001). Functional fitness did not mediate the relationship between physical activity and life satisfaction. Promoting physical activity may be useful for increasing older adults' functional fitness and life satisfaction, but the effects on functional fitness only occurred at a high level of physical activity, and the effect of physical activity on life satisfaction was not mediated by functional fitness.

Endoplasmic Reticulum Stress Mediates Renal Tubular Vacuolation in BK Polyomavirus-Associated Nephropathy.

Objective: This study aimed to explore the molecular mechanism of cytoplasmic vacuolation caused by BK polyomavirus (BKPyV) and thus search for potential target for drug repurposing. Methods: Morphological features of BK polyomavirus-associated nephropathy (BKPyVAN) were studied under light and electron microscopes. Microarray datasets GSE75693, GSE47199, and GSE72925 were integrated by ComBat, and differentially expressed genes (DEGs) were analyzed using limma. Furthermore, the endoplasmic reticulum (ER)-related genes obtained from GenCLiP 2.0 were intersected with DEGs. GO and KEGG enrichment pathways were performed with intersection genes by R package clusterProfiler. The single-cell RNA sequencing (scRNA-seq) from a BKPyVAN recipient was analyzed with a dataset (GSE140989) downloaded from Gene Expression Omnibus (GEO) as control for gene set variation analysis (GSVA). Immunohistochemistry and electron microscopy of kidney sections from drug-induced ERS mouse models were performed to explore the association of ERS and renal tubular vacuolation. Protein-protein interaction (PPI) network of the intersection genes was constructed to identify hub target. AutoDock was used to screen Food and Drug Administration (FDA)-approved drugs that potentially targeted hub gene. Results: Light and electron microscopes exhibited obvious intranuclear inclusions, vacuoles, and virus particles in BKPyV-infected renal tubular cells. Transcriptome analysis revealed 629 DEGs between samples of BKPyVAN and stable transplanted kidneys, of which 16 were ER-associated genes. GO analysis with the intersection genes illustrated that ERS-related pathways were significantly involved, and KEGG analysis showed a prominent enrichment of MAPK, Toll-like receptor, and chemokine signaling pathways. GSVA analysis of the proximal tubule revealed similar pathways enrichment. An electron microscope image of the kidney from ERS mouse models showed an obvious renal tubular vacuolation with prominent activation of ERS markers verified by immunohistochemistry. Furthermore, DDIT3 was identified as the hub gene based on PPI analysis, and ZINCOOOOO1531009 (Risedronate) was indicated to be a potential drug for DDIT3. Conclusion: ERS was involved in renal tubular cytoplasmic vacuolation in BKPyVAN recipients. Risedronate was screened as a potential drug for BKPyVAN by targeting DDIT3.

Paeoniflorin alleviates inflammatory response in IBS-D mouse model via downregulation of the NLRP3 inflammasome pathway with involvement of miR-29a.

Paeoniflorin has been traditionally used to treat pain and immunologic derangement in China. However, its detailed mechanism remains to be illuminated. We investigated the mechanism by which paeoniflorin alleviates the inflammatory response in a mouse model of irritable bowel syndrome with predominant diarrhea (IBS-D). C57BL/6 wild type (WT) and miR-29a knockout (KO) mice were randomly divided into control, model, rifaximin, and paeoniflorin groups (n = 7). IBS-D model was induced by single intracolonic instillation of 0.1 mL trinitro-benzene-sulfonic acid (TNBS, 50 mg/mL) combined with restraint stress for seven consecutive days. The treatment groups received rifaximin (100 mg/kg) and paeoniflorin (50 mg/kg) via intragastric administration for seven days, respectively. The results showed that the fecal water content, fecal pellet output, visceral sensitivity, and histopathological score after paeoniflorin treatment were lower than those of the model group in both WT and miR-29a KO mice (P < 0.05). In both lineage mice, damage was observed in the colon tissues of model group, while paeoniflorin treatment partially ameliorated the tissue damage. Serum levels of DAO, DLA, IL-1ß, IL-18, TNF-α, and MPO were decreased after paeoniflorin treatment (P < 0.05), with miR-29a KO mice in a lower level compared with that of WT mice. RT-PCR showed that the relative expression of miR-29a, NF-κB (p65), NLRP3, ASC, caspase-1, IL-1ß, and TNF-α was downregulated while NKRF was upregulated after paeoniflorin treatment (P < 0.05). Immunohistochemistry showed that intestinal epithelial protein levels of NLRP3, ASC, and caspase-1 decreased while those of Claudin-1 and ZO-1 increased in the paeoniflorin treatment group (P < 0.05). In general, compared with WT mice, NLRP3 inflammasome pathway targets was in much lower expression level than miR-29a KO mice. In conclusion, paeoniflorin could inhibit abnormal activation of the NLRP3 inflammasome pathway by inhibiting miR-29a in IBS-D, thereby relieving the inflammatory response of the intestinal mucosa and reconstructing the intestinal epithelial barrier.

Association of Physical Fitness Performance Tests and Anthropometric Indices in Taiwanese Adults.

BACKGROUND: The association between physical fitness performance tests and anthropometric indices is not clear. The study aims to explore the association between physical fitness performance and anthropometric indices in Taiwanese community-dwelling adults. This may help in monitoring anthropometric indices to improve physical fitness. METHODS: We recruited 2216 participants aged 23-64 years between 2014 and 2017. Physical fitness performance, including abdominal muscular endurance (60-s sit-up test), flexibility (sit-and-reach test), and cardiorespiratory endurance (3-min step test), was evaluated in all participants. The association of the physical fitness performance and anthropometric indices, including body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR), was analyzed using linear regression, with adjustments for age and gender. RESULTS: Body mass index was negatively associated with abdominal muscular endurance (p < 0.001) and cardiorespiratory endurance (p < 0.001). Neither BMI, WC, WHR, nor WHtR were significantly associated with flexibility. Abdominal muscle endurance, flexibility, and cardiorespiratory endurance were significantly lower in obese participants when obesity was defined using a BMI of ≥27, 30, and 35 kg/m2. Participants with central obesity that was defined as WC ≥ 90 cm in men and 80 cm in women and WHtR ≥ 0.6 had lower abdominal muscular endurance than those without central obesity. CONCLUSION: Body mass index is associated with abdominal muscular endurance and cardiorespiratory endurance in a reverse J-shaped manner. None of the anthropometric indices are significantly associated with flexibility. Obesity defined by BMI is linked to worse physical fitness performance and obesity defined using WHtR is linked to lower abdominal muscular endurance in Taiwanese community-dwelling adults.

The Relationships between Physical Activity and Life Satisfaction and Happiness among Young, Middle-Aged, and Older Adults.

Physical activity has benefits on physical and psychological health. The aims of this study were to investigate (1) the relationships between physical activity and life satisfaction and happiness in young, middle-aged, and older adults while controlling for demographic characteristics, and (2) the relationships between age and life satisfaction and happiness for different physical activity levels. A total of 2345 healthy adults were recruited. Demographic characteristic, physical activity, life satisfaction, and happiness were collected. Participants were divided into young, middle-aged, and older adult groups based on age, and physical activity was categorized as high, moderate, and low. After controlling for demographic characteristics, participants with high and moderate activity levels had significantly higher life satisfaction and happiness than those with a low activity level across the total population and the three age groups. Age squared was a significant predictor of a positive curvilinear between age and life satisfaction and happiness. Physical activity was significantly related to life satisfaction and happiness in young, middle-aged, and older adults. In addition, life satisfaction and happiness increased with increasing age. The results support the promotion of physical activity.

Non-invasive urinary sediment double-immunostaining predicts BK polyomavirus associated-nephropathy in kidney transplant recipients.

BACKGROUND: The positive predictive value (PPV) of urinary decoy cells for diagnosing BK polyomavirus associated-nephropathy (BKPyVAN) is low. This study was designed to increase the PPV of urinary decoy cells for diagnosing BKPyVAN in kidney transplant recipients. METHODS: A total of 105 urine sediment samples from 105 patients with positive BK viruria and decoy cells were evaluated by automatic double-immunostaining with anti-HGD (a renal tubular marker) antibody + anti-SV40-T antibody or anti-S100P (an urothelial marker) antibody + anti-SV40-T antibody. RESULTS: Of the 105 patients, 76 (72.4%) had both HGD(+)/SV40-T(+) cells and S100P(+)/SV40-T(+) cells (group A), 24 (22.9%) had only S100P(+)/SV40-T(+) cells (group B), and 5 (4.6%) had only S100P(-)/HGD(-)/SV40-T(+) cells (group C). Seventy patients in group A (92.1%), 3 patients in group B (12.5%), and no patients in group C were diagnosed with BKPyVAN. The area under the ROC curve of predicting BKPyVAN by decoy cells was 0.531 (0.431-0.630), with an optimal cut-off value of 29 (per 10 high power field), a sensitivity of 45.8% (95% CI: 34.0-58.0%), and a specificity of 68.8% (95% CI: 50.0-83.9%). Besides, the area under the ROC curve of predicting BKPyVAN by plasma BKPyV load was 0.735 (95% CI: 0.632-0.822), with an optimal cut-off value of 1,000 copies/mL, a sensitivity of 61.1% (95% CI: 48.9-72.4%) and a specificity of 84.2% (95% CI: 60.4-96.6%). In contrast, the PPV, negative predictive value, sensitivity, and specificity of HGD(+)/SV40-T(+) cells for diagnosing BKPyVAN were 92.1% [95% confidence interval (CI): 83.0-96.7%], 89.7% (95% CI: 71.5-97.3%), 95.9% (95% CI: 87.7-98.9%), and 81.3% (95% CI: 63.0-92.1%) respectively. CONCLUSIONS: Double-immunostaining with anti-HGD or anti-S100P and anti-SV40-T antibodies helps to identify the origin of decoy cells and diagnose BKPyVAN.

Pathological characteristics of BK polyomavirus-associated nephropathy with glomerular involvement.

BACKGROUND: This study aimed to investigate the pathological characteristics of BK polyomavirus (BKPyV)-associated nephropathy (BKPyVAN) with glomerular involvement in kidney transplant recipients. METHODS: Forty-four patients with glomerular BKPyV infection were retrospectively included for analysis. Immunohistochemical (IHC) staining was performed on paraffin sections using monoclonal mouse anti-SV40 large T antigen antibody. RESULTS: In BKPyV-infected glomeruli, the glomerular parietal epithelial cells (GPECs) were swollen, hyperchromatic, and enlarged, with an increased nuclear to cytoplasm (N/C) ratio and smudgy basophilic intra-nuclear viral inclusions. IHC staining revealed the distribution of BKPyV involvement in GPECs, podocytes, and shedding cells within Bowman's space. Notably, BKPyV affected GPEC proliferation and caused crescent formation (7 biopsies, 15.9%). Three biopsies exhibited fibrous crescents and the absence of viral inclusions. The other 4 biopsies exhibited cellular and fibro-cellular crescents, with viral cytopathic changes and positive IHC staining in the proliferative GPECs. Electron microscopy showed viral particles in both GPECs and podocytes. BKPyV-infected GPECs were degenerative, with mitochondrial swelling, endoplasmic reticulum expansion, and multi-layered membranous structure formation. Twelve (27.3%) patients received repeat biopsies within 1.6 to 39.5 months (median: 13.5 months), but none revealed persistent glomerular BKPyV infection. CONCLUSIONS: Distinct glomerular changes in BKPyVAN biopsies should raise the possibility of glomerular involvement.

Detection of Proximal Tubule Involvement by BK Polyomavirus in Kidney Transplant Recipients With Urinary Sediment Double-Immunostaining.

Background: The extent and depth of BK polyomavirus (BKPyV) infection in renal allograft correlate with prognosis. This study was designed to evaluate the value of urinary sediment double-immunostaining for predicting BKPyV infection in proximal tubular epithelium. Materials and methods: A total of 76 urine sediment cell blocks, as well as the corresponding transplanted kidney tissues with BK polyomavirus associated-nephropathy (BKPyVAN), were evaluated by automatic double-immunostaining with anti-58-kDa Golgi protein (58K, a proximal renal tubular marker) + anti-SV40-T and anti-homogentisate 1, 2-dioxygenase (HGD, a renal tubular marker) + anti-SV40-T. Results: Immunohistochemical staining demonstrated that 58K was expressed in proximal tubular epithelium but not in distal tubular epithelium or transitional epithelium. Of the 76 patients, 28 (36.8%) had urinary 58K(+)/SV40-T(+) cells and HGD(+)/SV40-T(+) cells, 41 (53.9%) had only HGD(+)/SV40-T(+) cells, one (1.3%) had only 58K(+)/SV40-T(+) cells, and six (7.9%) had only 58K(-)/HGD(-)/SV40-T(+) cells. The presence of urinary 58K(+)/SV40-T(+) cells was correlated with BKPyV infection in proximal tubular epithelium (P < 0.001, r = 0.806). The mean extent of SV40-T staining was significantly more extensive in patients with urinary 58K(+)/SV40-T(+) cells than those without urinary 58K(+)/SV40-T(+) cells (21.4 vs. 12.0%, P < 0.001). The positive predictive value, negative predictive value, sensitivity, and specificity of urinary 58K(+)/SV40-T(+) cells for predicting BKPyV infection in proximal tubular epithelium were 89.7% (95% CI: 71.5-97.3%), 91.5% (95% CI: 78.7-97.2%), 86.7% (95% CI: 68.4-95.6%), and 93.5% (95% CI: 81.1-98.3%), respectively. Conclusion: Urinary sediment double-immunostaining with anti-58K and anti-SV40-T is valuable for predicting the extent and depth of BKPyV infection in renal allograft.