Infection Control Experiences and Educational Needs of Geriatric Care Workers in Long-Term Care Facilities: A Pilot Study.

BACKGROUND: In the post-COVID-19 condition, infection control education is important for geriatric care workers who care for the elderly and are vulnerable to emerging infectious diseases. This study was conducted to enhance the insight into the experiences of geriatric care workers in managing novel infectious diseases (COVID-19) and to identify the newly required educational requirements necessary to effectively implement infectious disease control. METHODS: This is a qualitative and pilot study using focus group interviews. Data from 10 participants were collected using a focus group interview. The data were analyzed using Qualitative content analysis. RESULTS: The findings showed that geriatric healthcare workers experienced difficulties following infection control protocols and emotional distress related to visitor restrictions and had an increased workload. The participants requested further education regarding general knowledge of infectious diseases to decrease their fears of infection and reported that visual and practical teaching methods were preferable. CONCLUSIONS: Further attention is needed regarding the education of infection control to strengthen infection prevention in long-term care facilities vulnerable to the spread of emerging infectious diseases.

Evolution of Cell-Type-Specific RNA Aptamers via Live Cell-Based SELEX.

Live cell-based SELEX (Systematic Evolution of Ligand EXponential enrichment) is a promising approach for identifying aptamers that can selectively bind to a cell-surface receptor or recognize a particular target cell population. In particular, it offers a facile selection strategy for some special cell-surface proteins that are originally glycosylated or heavily posttranslationally modified and are unavailable in their native/active conformation after in vitro expression and purification. In this chapter, we describe a generalized procedure for evolution of cell type-specific RNA aptamers targeting a cell membrane bound target by combining the live cell-based SELEX strategy with high-throughput sequencing (HTS) and bioinformatics analysis.

Development of a Tool to Measure Compliance with Infection Prevention Activities Against Emerging Respiratory Infectious Diseases among Nurses Working in Acute Care and Geriatric Hospitals.

BACKGROUND: This study developed a preliminary instrument to measure nurses' infection prevention compliance against emerging respiratory infectious diseases and to verify the reliability and validity of the developed instrument. METHOD: The participants were 199 nurses working at a university hospital with more than 800 beds and two long-term care hospitals. Data were collected in May 2022. RESULTS: The final version of the developed instrument consisted of six factors and 34 items, with an explanatory power of 61.68%. The six factors were equipment and environment management and education, hand hygiene and respiratory etiquette, infection risk assessment and flow management, protection of employees in contact with infected patients, ward access management of patients with infectious diseases, and wearing and removing personal protective equipment. We verified the convergent and discriminant validities of these factors. The instrument's internal consistency was adequate (Cronbach's α=0.82), and the Cronbach's α of each factor ranged from 0.71 to 0.91. CONCLUSION: This instrument can be utilized to determine the level of nurses' compliance with infection prevention activity against emerging respiratory infectious diseases and will contribute to measuring the effectiveness of future programs promoting infection-preventive activities.

Factors Influencing Frontline Screening Nurses' Depression as a Long-Term Impact of COVID-19.

Frontline screening nurses experienced exhaustion and depressive symptoms as a long-term impact of COVID-19. This study aimed to explore fatigue, depression, and empowerment among frontline screening nurses and examine the factors influencing depression. This was a descriptive cross-sectional study. The study included 140 frontline screening nurses in South Korea recruited from February to March 2021. The measures included a fatigue scale, the Text of Items Measuring Empowerment (TIME), and the Center for Epidemiological Studies Depression Scale (CES-D). The STROBE checklist was used for reporting aspects of the cross-sectional design. Frontline screening nurses showed high fatigue scores (M = 3.47, SD = 0.55), and 55.7% (n = 78) of them were depressed and had low empowerment scores (M = 3.53, SD = 0.69). Empowerment and fatigue were predictors of depression. Increased empowerment and decreased fatigue were important in decreasing depression. Therefore, efforts to provide sufficient staffing, screening for depression, and listening to nurses' voices are necessary.

The Effect of Dicer Knockout on RNA Interference Using Various Dicer Substrate Small Interfering RNA (DsiRNA) Structures.

Small interfering RNAs (siRNAs) are artificial molecules used to silence genes of interest through the RNA interference (RNAi) pathway, mediated by the endoribonuclease Dicer. Dicer-substrate small interfering RNAs (DsiRNAs) are an alternative to conventional 21-mer siRNAs, with an increased effectiveness of up to 100-fold compared to traditional 21-mer designs. DsiRNAs have a novel asymmetric design that allows them to be processed by Dicer into the desired conventional siRNAs. DsiRNAs are a useful tool for sequence-specific gene silencing, but the molecular mechanism underlying their increased efficacy is not precisely understood. In this study, to gain a deeper understanding of Dicer function in DsiRNAs, we designed nicked DsiRNAs with and without tetra-loops to target a specific mRNA sequence, established a Dicer knockout in the HCT116 cell line, and analyzed the efficacy of various DsiRNAs on RNAi-mediated gene silencing activity. The gene silencing activity of all DsiRNAs was reduced in Dicer knockout cells. We demonstrated that tetra-looped DsiRNAs exhibited increased efficacy for gene silencing, which was mediated by Dicer protein. Thus, this study improves our understanding of Dicer function, a key component of RNAi silencing, which will inform RNAi research and applications.

Modifications of Rect-Spring to Enhance the Engagement of Ectopically Entrapped Molars with 2 Case Reports.

The Rect-spring appliance, used for the management of ectopically erupting molars, shows weak retention on mesially tilted molars. We present three modifications of the appliance for better engagement and their advantages. We describe cases of two 7-year-old patients with ectopically erupting maxillary first molars with a 2.2 mm and 2.5 mm depth of entrapment, respectively. The modified Rect-spring (mRS) was inserted between the ectopically erupting first molar and adjacent primary second molar, and exerted a distalization force with an interproximal wedging effect at the same time. After 3 months, the ectopically erupting first molars were successfully brought into proper occlusion. No discomfort was reported. The mRS is suitable for various locking cases except for severely tilted molars without requiring any laboratory procedures. We suggest it as the first choice for unlocking the first molars.

Comparing two health literacy measurements used for assessing older adults' medication adherence.

AIMS AND OBJECTIVES: To compare two health literacy measurements' ability to assess older adults' medication adherence by using the Korean Health Literacy Screening Questions (KHLSQ) and the Modified Korean Functional Health Literacy Test (M-KFHLT), and to identify an appropriate health literacy measurement. BACKGROUND: Lower health literacy has been associated with poorer medication adherence. Thus, health professionals should evaluate the available health literacy assessment instruments they are using and choose an appropriate instrument to assess health literacy to increase older adults' medication adherence. DESIGN: Following the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist, a descriptive, cross-sectional study was conducted. METHODS: From November 2017-May 2018, 116 community-dwelling older adults were recruited in South Korea. Questionnaires were completed during a face-to-face interview with each participant in a private room; health literacy was assessed using the KHLSQ and the M-KFHLT and medication adherence using the Morisky Medication Adherence Scale. RESULTS: Health literacy assessed using the KHLSQ was found to be a predictor of medication adherence, but was not a predictor when measured by the M-KFHLT. Having low income, multiple chronic diseases and vision problems were also significant factors related to medication adherence. CONCLUSIONS: This study suggests that health literacy was negatively associated with medication adherence. Furthermore, KHLSQ is an appropriate tool for healthcare providers to use when assessing health literacy to predict older adults' medication adherence. RELEVANCE TO CLINICAL PRACTICE: This finding indicated that healthcare providers should select an appropriate health literacy measurement that suits their purposes and the population they serve, particularly for older adults.

Molecular mechanisms of Dicer: endonuclease and enzymatic activity.

The enzyme Dicer is best known for its role as a riboendonuclease in the small RNA pathway. In this canonical role, Dicer is a critical regulator of the biogenesis of microRNA and small interfering RNA, as well as a growing number of additional small RNAs derived from various sources. Emerging evidence demonstrates that Dicer's endonuclease role extends beyond the generation of small RNAs; it is also involved in processing additional endogenous and exogenous substrates, and is becoming increasingly implicated in regulating a variety of other cellular processes, outside of its endonuclease function. This review will describe the canonical and newly identified functions of Dicer.

The anti-miR21 antagomir, a therapeutic tool for colorectal cancer, has a potential synergistic effect by perturbing an angiogenesis-associated miR30.

Colon cancer has the third highest incidence and mortality among cancers in the United States. MicroRNA-21 (miR21) has been described as an oncomir that is highly overexpressed in tumor tissue from colorectal cancer. Recent studies showed that silencing of miR21 through use of a miR21 inhibitor (anti-miR21) affected viability, apoptosis and the cell cycle in colon cancer cells. We identified an anti-miR21 that targets miR21 to inhibit genes by both post-transcriptional gene silencing and transcriptional gene silencing in the cytoplasm and nucleus, respectively. Overexpression of anti-miR21 in colon cancer cells caused changes in miRNA expression levels. We found that treatment with anti-miR21 down-regulated expression of miR30, which is involved in angiogenesis. In an in vitro angiogenesis assay, network formation induced by an angiogenesis activator was reduced upon treatment with anti-miR21. Sequence analysis of anti-miR21 and pri-miR30 revealed homology between anti-miR21 and the 3' end of pri-miR30, suggesting that anti-miR21 may bind to pri-miR30 and block processing of the miRNA processing. These results suggest anti-miR21 has a role not only in tumor growth but also in angiogenesis. Therefore, treatment with the anti-miR21 antagomir may have a synergistic effect mediated through suppression of miR30.

Efficient targeting and tumor retardation effect of pancreatic adenocarcinoma up-regulated factor (PAUF)-specific RNA replacement in pancreatic cancer mouse model.

The soluble protein pancreatic adenocarcinoma up-regulated factor (PAUF) plays an important role in pancreatic tumor progression and has begun to attract attention as a therapeutic target for pancreatic cancer. We herein present PAUF RNA-targeting gene therapy strategies with both targeting and therapeutic function using trans-splicing ribozyme (TSR) in pancreatic cancer. We developed adenoviral PAUF-targeting TSR (Rz) containing a PAUF-specific internal guide sequence (IGS) determined by library screening. This Rz harbors suicide gene, herpes simplex virus thymidine kinase (HSV-tk) or firefly luciferase (Luc) as a transgene for 3' exon replacement of PAUF RNAs. Ad-Rz-TK, Rz harboring the HSV-tk, showed significant inhibition of tumor growth in vivo as well as PAUF-dependent cell death in vitro via a successful trans-splicing reaction. Selective induction of Rz-controlled transgene in PAUF-expressing pancreatic cancer was confirmed through noninvasive in vivo imaging; a luminescence signal from Rz harboring Luc (Ad-Rz-Luc) was detectable only in pancreatic tumor sites, not in normal mice. In addition, a [(125)I] FIAU signal reflecting thymidine kinase expression through SPECT and ex vivo biodistribution was co-localized with the tumor sites when we treated with Ad-Rz-TK in orthotopic xenograft model. Taken together, these results imply that PAUF-targeting TSR can contribute to successful targeted gene therapy for pancreatic cancer.

Effects of education in an obesity control program for obese homemakers on body fat and flexibility in Korea.

The study aims to investigate the effects of education in an obesity-control program (EOCP) on the percentage of body fat and flexibility in obese women in Korea. Women with over 30% body fat were offered EOCP between July 2012 and October 2012. The EOCP consisted of an educational program, exercise program, and counseling. The numbers of participants both in the EOCP and control group were 17. The study was analyzed by Wilcoxon signed-rank test using the Statistical Analysis System package. The EOCP participants presented statistically significant increases in the degree of forward trunk flexion, but only the percentage of the body fat showed differences within the EOCP group. The EOCP improved flexibility in obese women, and can be used in local obesity-control programs.

Effects of muscular and aqua aerobic combined exercise on metabolic indices in elderly women with metabolic syndrome.

The purpose of this study was to investigate the effects of muscle strengthening exercise using elastic thera-band and aquatic aerobic combined exercise on metabolic syndrome index in elderly with metabolic syndrome. Fifty-four were assigned to muscle strengthening exercise group (n = 19), aquatic aerobic exercise group (n = 19), and combined exercise group (n = 16). The muscle strength exercise, aquatic aerobic exercise and combined exercise were provided three times a week for 12 weeks. Metabolic syndrome indices[Fasting blood glucose, triglyceride, high density lipoprotein cholesterol (HDL-C), systolic blood pressure, diastolic blood pressure and waist circumference] were measured before and after the program. One-way ANOVA, paired t-test and two-way repeated ANOVA were used with the SPSS program for data analysis. There was a significant difference in triglyceride (p < .001), HDL-C (p = .010) and waist circumference (p = .016). Triglyceride and waist circumference was significantly decreased in combined group than muscle strength exercise group and aquatic exercise group. HDL-C was significantly increased in combined group than muscle strength exercise group. The results indicate that combined exercise was more effective in the improvement of dyslipidemia and abdominal obesity.

Effects of nordic walking on body composition, muscle strength, and lipid profile in elderly women.

PURPOSE: The purpose of this study was to investigate the effects of Nordic walking on body composition, muscle strength, and lipid profile in elderly women. METHOD: Sixty-seven women were assigned to the Nordic walking group (n = 21), the normal walking group (n = 21), and the control group (n = 25). Nordic walking and normal walking were performed three times a week for 12 weeks. Body weight, body mass index, total body water, skeletal muscle mass, percent body fat, grip strength, sit to stand, arm curl, total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were measured before and after the program. A Chi-square test, one way analysis of variance, paired t test and repeated-measure two-factor analysis were used with the SAS program for data analysis. RESULTS: There was a significant difference in the weight (F=8.07, p<.001), grip strength (F=10.30, p<.001), sit to stand (F=16.84, p<.001), arm curl (F=41.16, p<.001), and total cholesterol (F=5.14, p=.009) measurements between the groups. In addition, arm curl was significantly increased in the Nordic walking group compared to the normal walking group and the control group. CONCLUSION: The results indicate that Nordic walking was more effective than normal walking in improving upper extremity strength.

Deep Sequencing Analyses of DsiRNAs Reveal the Influence of 3' Terminal Overhangs on Dicing Polarity, Strand Selectivity, and RNA Editing of siRNAs.

25/27 Base duplex RNAs that are substrates for Dicer have been demonstrated to enhance RNA interference (RNAi) potency and efficacy. Since the target sites are not always equally susceptible to suppression by small interfering RNA (siRNA), not all 27-mer duplexes that are processed into the corresponding conventional siRNAs show increased potency. Thus random designing of Dicer-substrate siRNAs (DsiRNAs) may generate siRNAs with poor RNAi due to unpredictable Dicer processing. Previous studies have demonstrated that the 3'-overhang affects dicing cleavage site and the orientation of Dicer entry. Moreover, an asymmetric 27-mer duplex having a 3' two-nucleotide overhang and 3'-DNA residues on the blunt end has been rationally designed to obtain greater efficacy. This asymmetric structure directs dicing to predictably yield a single primary cleavage product. In the present study, we analyzed the in vitro and intracellular dicing patterns of chemically synthesized duplex RNAs with different 3'-overhangs. Consistent with previous studies, we observed that Dicer preferentially processes these RNAs at a site 21-22 nucleotide (nt) from the two-base 3'-overhangs. We also observed that the direction and ability of human Dicer to generate siRNAs can be partially or completely blocked by DNA residues at the 3'-termimi. To examine the effects of various 3'-end modifications on Dicer processing in cells, we employed Illumina Deep sequencing analyses to unravel the fates of the asymmetric 27-mer duplexes. To validate the strand selection process and knockdown capabilities we also conducted dual-luciferase psiCHECK reporter assays to monitor the RNAi potencies of both the "sense" (S) and "antisense" (AS) strands derived from these DsiRNAs. Consistent with our in vitro Dicer assays, the asymmetric duplexes were predictably processed into desired primary cleavage products of 21-22-mers in cells. We also observed the trimming of the 3' end, especially when DNA residues were incorporated into the overhangs and this trimming ultimately influenced the Dicer-cleavage site and RNAi potency. Moreover, the observation that the most efficacious strand was the most abundant revealed that the relative frequencies of each "S" or "AS" strand are highly correlated with the silencing activity and strand selectivity. Collectively, our data demonstrate that even though the only differences between a family of DsiRNAs was the 3' two-nuclotide overhang, dicing polarity and strand selectivity are distinct depending upon the sequence and chemical nature of this overhang. Thus, it is possible to predictably control dicing polarity and strand selectivity via simply changing the 3'-end overhangs without altering the original duplex sequence. These optimal design features of 3'-overhangs might provide a facile approach for rationally designing highly potent 25/27-mer DsiRNAs.

Signal amplification by magnetic force on polydiacetylene supramolecules for detection of prostate cancer.

A method in which a permanent magnet is introduced onto polydiacetylene (PDA) vesicle chips is introduced for enhancement of the fluorescence of PDA vesicles. This strategy can be applied to general antibody-based PDA vesicle chips to detect clinically important biomarkers for disease diagnosis.

A role for human Dicer in pre-RISC loading of siRNAs.

RNA interference is a powerful mechanism for sequence-specific inhibition of gene expression. It is widely known that small interfering RNAs (siRNAs) targeting the same region of a target-messenger RNA can have widely different efficacies. In efforts to better understand the siRNA features that influence knockdown efficiency, we analyzed siRNA interactions with a high-molecular weight complex in whole cell extracts prepared from two different cell lines. Using biochemical tools to study the nature of the complex, our results demonstrate that the primary siRNA-binding protein in the whole cell extracts is Dicer. We find that Dicer is capable of discriminating highly functional versus poorly functional siRNAs by recognizing the presence of 2-nt 3' overhangs and the thermodynamic properties of 2-4 bp on both ends of effective siRNAs. Our results suggest a role for Dicer in pre-selection of effective siRNAs for handoff to Ago2. This initial selection is reflective of the overall silencing potential of an siRNA.

Unilamellar nanosheet of layered manganese cobalt nickel oxide and its heterolayered film with polycations.

The exfoliation of layered Li[Mn(1/3)Co(1/3)Ni(1/3)]O(2) into individual monolayers could be achieved through the intercalation of quaternary tetramethylammonium (TMA(+)) ions into protonated metal oxide. An effective exfoliation occurred when the TMA(+)/H(+) ratio was 0.5-50. Reactions outside this range produced no colloidal suspension, but all the manganese cobalt nickel oxides precipitated. Atomic force microscopy and transmission electron microscopy clearly demonstrated that exfoliated manganese cobalt nickel oxide nanosheets have a nanometer-level thickness, underscoring the formation of unilamellar nanosheets. The maintenance of the hexagonal atomic arrangement of the manganese cobalt nickel oxide layer upon the exfoliation was confirmed by selected area electron diffraction analysis. According to diffuse reflectance ultraviolet--visible spectroscopy, the exfoliated manganese cobalt nickel oxides displayed distinct absorption peaks at approximately 354 and approximately 480 nm corresponding to the d-d transitions of octahedral metal ions, which contrasted with the featureless spectrum of the pristine metal oxide. In the light of zeta potential data showing the negative surface charge of manganese cobalt nickel oxide nanosheets, a heterolayered film of manganese cobalt nickel oxide and conductive polymers could be prepared through the successive coating process with colloidal suspension and polycations. The UV--vis and X-ray diffraction studies verified the layer-by-layer ordered structure of the obtained heterolayered film, respectively.

Cancer cell targeting with mouse TERT-specific group I intron of Tetrahymena thermophila.

Telomerase reverse transcriptase (TERT), which prolongs the replicative life span of cells, is highly upregulated in 85-90% of human cancers, whereas most normal somatic tissues in humans express limited levels of the telomerase activity. Therefore, TERT has been a potential target for anticancer therapy. Recently, we described a new approach to human cancer gene therapy, which is based on the group I intron of Tetrahymena thermophila. This ribozyme can specifically mediate RNA replacement of human TERT (hTERT) transcript with a new transcript harboring anticancer activity through a trans-splicing reaction, resulting in selective regression of hTERT-positive cancer cells. However, to validate the therapeutic potential of the ribozyme in animal models, ribozymes targeting inherent transcripts of the animal should be developed. In this study, we developed a Tetrahymena-based trans-splicing ribozyme that can specifically target and replace the mouse TERT (mTERT) RNA. This ribozyme can trigger transgene activity not only also in mTERT-expressing cells but hTERT-positive cancer cells. Importantly, the ribozyme could selectively induce activity of the suicide gene, a herpes simplex virus thymidine kinase gene, in cancer cells expressing the TERT RNA and thereby specifically hamper the survival of these cells when treated with ganciclovir. The mTERT-targeting ribozyme will be useful for evaluation of the RNA replacement approach as a cancer gene therapeutic tool in the mouse model with syngeneic tumors.

RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein.

Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of a newly emerged disease SARS. The SARS-CoV nucleocapsid (N) protein is one of the most abundant structural proteins and serves as a diagnostic marker for accurate and sensitive detection of the virus. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant N protein, we selected a high-affinity RNA aptamer capable of binding to N protein with a dissociation constant of 1.65 nM. Electrophoretic mobility shift assays and RNA competition experiments showed that the selected aptamer recognized selectively the C-terminal region of N protein with high specificity. Using a chemiluminescence immunosorbent assay and a nanoarray aptamer chip with the selected aptamer as an antigen-capturing agent, we could sensitively detect N protein at a concentration as low as 2 pg/ml. These aptamer-antibody hybrid immunoassays may be useful for rapid, sensitive detection of SARS-CoV N protein.