Micropreparative Gel Electrophoresis for Purification of Nanoscale Bioconjugates.

Conventional techniques for purifying macromolecular conjugates often require complex and costly installments that are inaccessible to most laboratories. In this work, we develop a one-step micropreparative method based on a trilayered polyacrylamide gel electrophoresis (MP-PAGE) setup to purify biological samples, synthetic nanoparticles, as well as biohybrid complexes. We apply this method to recover DNA from a ladder mixture with yields of up to 90%, compared to the 58% yield obtained using the conventional crush-and-soak method. MP-PAGE was also able to isolate enhanced yellow fluorescence protein (EYFP) from crude cell extract with 90% purity, which is comparable to purities achieved through a more complex two-step purification procedure involving size exclusion and immobilized metal-ion affinity chromatography. This technique was further extended to demonstrate size-dependent separation of a commercial mixture of graphene quantum dots (GQDs) into three different fractions with distinct optical properties. Finally, MP-PAGE was used to isolate DNA-EYFP and DNA-GQD bioconjugates from their reaction mixture of DNA and EYFP and GQD precursors, samples that otherwise could not be effectively purified by conventional chromatography. MP-PAGE thus offers a rapid and versatile means of purifying biological and synthetic nanomaterials without the need for specialized equipment.

Treatment and mortality risk of older adults with non-small cell cancer in Taiwan: A population-based cohort study.

BACKGROUND: Older patients tend to have decreased physical functions and more comorbidities than younger patients. At present, the best management for very elderly patients with lung cancer is not known. In this study, we aimed to investigate treatment and mortality risk of older adults with non-small cell cancer (NSCLC) in Taiwan. METHODS: This study analyzed data from the Taiwan Cancer Registry database. Patients aged ≥80 years with newly diagnosed NSCLC between 2010 and 2017 were included. Treatment options were categorized as curative, palliative, and no treatment. Patients were followed up until death or December 31, 2020. Univariable and multivariable Cox proportional hazards models were used to estimate mortality risk, and Kaplan-Meier survival curves were drawn. RESULTS: A total of 11 941 patients, aged ≥80 years, with newly diagnosed NSCLC between 2010 and 2017 were identified from the Taiwan Cancer Registry and followed up until 2020. The mean age was 84.4 ± 3.7 years old, and 7468 (62.54%) were men. The Kaplan-Meier survival curves showed significant differences across the three treatment options (log-rank p < 0.001). Results from multivariate Cox regression demonstrated that patients on palliative treatment (adjusted HR: 0.52, 95% CI: 0.48-0.56, p < 0.001) and curative treatment (adjusted HR: 0.45, 95% CI: 0.42-0.48, p < 0.001) had a significantly lower mortality risk than those with no treatment. The subgroup analyses stratified by cancer stages also showed consistent findings. CONCLUSION: Elderly patients with NSCLC had significantly decreased mortality risk when receiving curative or palliative treatment compared with those without treatment. In the future, further studies are warranted to investigate complications and quality of life of elderly patients with NSCLC during palliative or curative treatment.

Phenome-wide analysis of Taiwan Biobank reveals novel glycemia-related loci and genetic risks for diabetes.

To explore the complex genetic architecture of common diseases and traits, we conducted comprehensive PheWAS of ten diseases and 34 quantitative traits in the community-based Taiwan Biobank (TWB). We identified 995 significantly associated loci with 135 novel loci specific to Taiwanese population. Further analyses highlighted the genetic pleiotropy of loci related to complex disease and associated quantitative traits. Extensive analysis on glycaemic phenotypes (T2D, fasting glucose and HbA1c) was performed and identified 115 significant loci with four novel genetic variants (HACL1, RAD21, ASH1L and GAK). Transcriptomics data also strengthen the relevancy of the findings to metabolic disorders, thus contributing to better understanding of pathogenesis. In addition, genetic risk scores are constructed and validated for absolute risks prediction of T2D in Taiwanese population. In conclusion, our data-driven approach without a priori hypothesis is useful for novel gene discovery and validation on top of disease risk prediction for unique non-European population.

Effects of Tongue-Strengthening Exercise on Tongue Strength Reserve and Detraining Effects among Healthy Adults: A Randomized Controlled Trial.

INTRODUCTION: Tongue strength reserve is the difference between the maximum isometric pressure (MIP) and swallowing pressure of the tongue. People with decreased tongue strength reserve may have a higher risk of presbyphagia or dysphagia. Thus, this study explored the effects of tongue strengthening exercise (TSE) on tongue strength reserve and detraining effects in healthy adults. MATERIALS AND METHODS: In total, 102 healthy volunteers without any reported history of speech or swallowing deficits were recruited and assigned to experimental (n = 50) and control groups (n = 52). Exercises in the experimental group consisted of compressing an air-filled bulb between the tongue and hard palate for 30 min a day, 5 days a week, for 8 weeks. Thereafter, the experimental group underwent a 4-week detraining period. RESULTS: Following the TSE training, posterior tongue strength reserve (F = 4.92, p = 0.029) of the experimental group was significantly higher than that of the control group. No significant detraining effects were observed on the MIP and swallowing pressure from 4 weeks after the completion of TSE training. CONCLUSIONS: According to the study results, TSE may be an effective approach for improving swallowing function.

[Preventive Care for Aspiration Pneumonia in Patients With an Indwelling Nasogastric Tube].

More than 190,000 patients have been fitted with an indwelling nasogastric tube in Taiwan. Nasogastric tube feeding is a strategy for temporary or long-term feeding and for preventing aspiration pneumonia in patients with dysphagia. However, aspiration pneumonia is a common complication in patients fitted with a nasogastric tube. Displacement of the nasogastric tube, inappropriate length of the placed nasogastric tube resulting gastroesophageal reflux, inappropriate feeding posture and speed, and excessive gastric residual volume resulting in vomiting are important risk factors associated with aspiration pneumonia. In this article, the literature and clinical practice experiences were reviewed to identify methods used to confirm the length and positioning of nasogastric tubes. We further explored strategies for preventing oral-care and tube-feeding-induced aspiration pneumonia during the period of indwelling nasogastric tube. These strategies included adjusting the patient's positions for receiving tube feedings, feeding techniques, methods and frequency of gastric residual volume measurement, and conditions for stopping nasogastric feeding. This exploration was intended to provide nursing staff with relevant knowledge and skills for preventing aspiration pneumonia in patients with indwelling nasogastric tube to improve patient safety.

Association between teeth loss and nasogastric tube feeding dependency in older adults from Taiwan: a retrospective cohort study.

BACKGROUND: To examine the association between teeth loss and nasogastric tube feeding dependency in older people. METHODS: The National Health Interview Survey (NHIS) 2005, 2009, and 2013 in Taiwan. Participants were selected by a multistage stratified sampling method and baseline characteristics, including socioeconomic status and health habits, were obtained by well-trained interviewers. The NHIS was linked with the National Health Insurance research database 2000-2016 and the National Deaths Dataset, which contains all the medical information of ambulatory and inpatient care. Cox regression was used to examine the association between the number of teeth lost and nasogastric tube feeding dependency. RESULTS: There were 6165 adults older than 65 years old enrolled in the analysis, with 2959 male (48%) and the mean (SD) age was 73.95(6.46) years old. The mean follow-up duration was 6.5(3.3) years. Regarding the teeth loss categories, 1660 (26.93%), 2123 (34.44%), and 2382 (38.64%) of participants were categorized as having no teeth loss, loss of 1-9 teeth, and loss of 10-28 teeth, respectively. During 39,962 person-years of follow-up, new-onset nasogastric feeding dependency was recognized in 220(13.25%), 256(12.06%), and 461(19.35%) participants who were categorized as having no teeth loss, loss of 1-9 teeth, and loss of 10-28 teeth, respectively. Kaplan-Meier curves demonstrated significant findings (Log-rank P < 0.01). After potential confounders were adjusted, compared with those without teeth loss, older adults who had lost 10-28 teeth had significantly increased risks of occurrence nasogastric feeding dependency (AHR, 1.31; 95% CI, 1.05-1.62; p-value = 0.02). Furthermore, a significant dose-response relation between the number of teeth lost and increased risk of nasogastric feeding was found (p for trend< 0.01). CONCLUSIONS: Older adults who had lost 10-28 teeth had a significantly increased risk of nasogastric tube feeding dependency. Early identification of the oral disease is crucial for the prevention of the occurrence of teeth loss and the following nutrition problems, which would reduce risk of nasogastric tube feeding dependency.

Migration and invasion of NSCLC suppressed by the downregulation of Src/focal adhesion kinase using single, double and tetra domain anti- CEACAM6 antibodies.

Carcinoembryonic antigen-related cell adhesion molecules 6 (CEACAM6) is a cell adhesion receptor. Expression of CEACAM6 in non-small cell lung cancer (NSCLC) associated with tumor progression and metastatic condition via Src/FAK signaling pathway. We established three anti-CEACAM6 antibodies with valences, which were designed to be monomeric sdAb, bivalent sdAb (2Ab), and tetravalent sdAb (4Ab). The anti-CEACAM6 antibodies can be used to target CEACAM6 overexpressing NSCLC. Anti-CEACAM6 antibodies, sdAb, 2Ab and 4Ab, were modified with different valency via protein engineering. sdAb and multivalent sdAbs (2Ab & 4Ab) were expressed and purified from E.coli and CHO cells, respectively. We compared the effect of anti-CEACAM6 antibodies with doxorubicin in NSCLC cell line both in vitro and in vivo. The 4Ab showed significant effect on cell viability. In addition, A549 cells treated with 2Ab and 4Ab inhibited the invasion and migration. In western blot, the 2Ab and 4Ab showed significant inhibition of phospho FAK domain Ty397 that is essential for activation of Src kinase family. Meanwhile, overall protein analysis revealed that 2Ab and 4Ab potently inhibited the phosphorylation of pSRC, pERK, pFAK, pAKT, MMP-2, MMP-9 and N-cadherin. Anti-tumor effect was observed in an A549 NSCLC xenograft model treated with 2Ab or 4Ab compared with doxorubicin. Confocal analysis showed higher targeting ability of 4Ab than that of 2Ab at 4 h incubation. Our data suggests that 2Ab and 4Ab inhibits EMT-mediated migration and invasion via suppression of Src/FAK signaling, which exhibits therapeutic efficiency for NSCLC treatment.

Source identification of HIV-1 transmission in three lawsuits Using Ultra-Deep pyrosequencing and phylogenetic analysis.

BACKGROUND/PURPOSE: Intentional transmission of HIV-1 is a crime. Identifying the source of transmission between HIV-1 infected cases using phylogenetic analysis has limitations, including delayed examinations after the initiation of infection and ambiguity of phyletic relationships. This study was the first to introduce phylogenetic tree Results as forensic evidence in a trial in Taiwan. METHODS: Three lawsuit cases from different district courts in Taiwan were chosen for this study. We identified the source of transmission between individuals in each lawsuit based on the maximum likelihood and Bayesian phylogenetic tree analyses using the HIV-1 sequences from molecular cloning and ultra-deep pyrosequencing (UDPS). Two gene regions of the HIV genome, env and gag, were involved. RESULTS: The results of phylogenetic analysis using sequences from molecular cloning were clear and evidential enough in lawsuits 1 and 3. Due to the delayed sampling time, the result of sequences from molecular cloning in lawsuit 2 was ambiguous. Combined with the analyzed result of sequences from UDPS and epidemiological information, the source of transmission in lawsuit 2 was further identified. CONCLUSION: Hence phylogenetic analyses cannot exclude the possibility of unsampled intermediaries, the data interpretation should be more careful and conservative, and it should not be considered as the only evidence for the source identification in a trial without epidemiological or serological information. The evaluation of the introduced UDPS method in the identification of transmission source has shown that the validity and evidential effects were still limited and need further optimization.

Analysis of Labial and Lingual Strength among Healthy Chinese Adults in Taiwan.

This study collected 11 parameters regarding the labial and lingual strength for maximum isometric and swallowing tasks among 150 healthy Chinese adults in Taiwan. Measurements were performed using the Iowa Oral Performance Instrument (IOPI). All of the labial and lingual strength parameters were measured three times. The maximal value of three trials represents the pressure of every parameter. The overall mean (±standard deviation) and maximum isometric pressures of the lips, anterior tongue, and posterior tongue were 24.81 ± 5.64, 55.95 ± 14.13, and 53.23 ± 12.24 kPa, respectively. The mean value of posterior tongue strength was less than that of the anterior tongue by approximately 5%. The percentages of maximum isometric tongue pressure during the swallowing of saliva and water were 85% and 80% for the anterior tongue and 90% and 81% for the posterior tongue, respectively. The average endurances for the anterior tongue and posterior tongue were 13.86 ± 7.08 and 10.06 ± 5.40 s, respectively. The maximum isometric pressures were greater than both the saliva and water swallowing pressures, and the saliva swallowing pressures were greater than the water swallowing pressures. A value of 33 kPa in maximum isometric pressure could serve as a demarcation of weak tongue strength for healthy Chinese adults. As for the repeated trials of labial and lingual strength, there were no statistically significant differences for any of the pressures obtained from the 11 labial and lingual strength parameters. The normative data can be used for the objective assessment of labial and lingual strength in healthy Chinese adults.

Evaluation of an Oral Hygiene Education Program for Staff Providing Long-Term Care Services: A Mixed Methods Study.

BACKGROUND: Oral hygiene is often neglected in clients receiving long-term care, suggesting that long-term care workers require formal oral hygiene education. Thus, the aim of this study was to investigate the effects of oral hygiene education on long-term care workers. METHODS: This study utilized a mixed methods design. Eighty long-term care workers were recruited for participation in the oral hygiene education program, which employed three teaching methods: narration with multimedia presentation, demonstration, and teach-back. The effect of the education program on the participants' level of oral hygiene knowledge, attitudes, and skills was measured using a structured questionnaire that was administered both pre- and post-delivery of the education program. Three months later, all participants submitted a self-report of their oral hygiene skills, and six participants completed a telephone interview. Quantitative data were analyzed using paired t-tests, and qualitative data were manually analyzed and coded. RESULTS: Scores of oral hygiene knowledge (p < 0.001), attitudes (p = 0.001), and oral cleaning daily frequency for clients (p < 0.001), were significantly higher three months after undertaking the educational program. CONCLUSIONS: This preliminary study suggests that oral hygiene education may be effective in improving oral hygiene knowledge, attitudes, and skills among long-term care staff.

Effects of an Eating Ability Promotion Program for Community-Dwelling Older Adults.

OBJECTIVE: Taiwan is projected to become a super-aged society in 2026. Taiwan's government introduced an innovative preventive care policy to improve aging-related conditions in 2017. In this study, we examine the effectiveness of an eating ability promotion program (EAPP) on the physical and mental performance of community-dwelling older adults. DESIGN: Single-masked, cluster randomized trial. SETTING: Two community care stations in Taichung, Taiwan. PARTICIPANTS: Volunteers were recruited from these 2 community care stations. The 70 participants were aged ≥60 years, able to walk independently and take care of themselves, and understood Mandarin or Taiwanese. INTERVENTION: The volunteers were randomly assigned to an intervention; 40 participated in EAPP training courses (experimental group) and 30 participated in originally scheduled activities only (control group). EAPP training courses were conducted in the care stations 4 hours per week for 12 weeks (for a total of 48 hours). MEASURES: The physical performance outcomes were oral health, nutritional status, and fragility, assessed using the Oral Health Assessment Tool, the Mini Nutritional Assessment, and the Study of Osteoporotic Fractures fragility index, respectively. Cognitive function was evaluated with the Mini-Cog test. Measurements were performed at baseline, at the end of the 12-week intervention, and 1 month later. RESULTS: Following the EAPP intervention, controlling for baseline differences, the oral health (F = 33.29, P < .001), nutritional status (F = 7.30, P = .009), and scale of fragility (F = 19.05, P < .001) of the participants in the experimental group were significantly better than those reported in the control group. CONCLUSIONS AND IMPLICATIONS: Results of this preliminary study suggest that the EAPP intervention may be an effective approach for improving oral health, nutritional status, and fragility in community-dwelling older adults. This training course, which provides clear and concise information regarding eating ability strategies, should undergo further evaluation and, if demonstrated to be effective and cost-effective in broader trials, may be useful in promoting healthy living.

Templating colloidal sieves for tuning nanotube surface interactions and optical sensor responses.

Surfactants offer a tunable approach for modulating the exposed surface area of a nanoparticle. They further present a scalable and cost-effective means for suspending single-walled carbon nanotubes (SWCNTs), which have demonstrated practical use as fluorescence sensors. Though surfactant suspensions show record quantum yields for SWCNTs in aqueous solutions, they lack the selectivity that is vital for optical sensing. We present a new method for controlling the selectivity of optical SWCNT sensors through colloidal templating of the exposed surface area. Colloidal nanotube sensors were obtained using various concentrations of sodium cholate, and their performances were compared to DNA-SWCNT optical sensors. Sensor responses were measured against a library of bioanalytes, including neurotransmitters, amino acids, and sugars. We report an intensity response towards dopamine and serotonin for all sodium cholate-suspended SWCNT concentrations. We further identify a selective, 14.1 nm and 10.3 nm wavelength red-shifting response to serotonin for SWCNTs suspended in 1.5 and 0.5 mM sodium cholate, respectively. Through controlled, adsorption-based tuning of the nanotube surface, this study demonstrates the applicability of sub-critical colloidal suspensions to achieve selectivities exceeding those previously reported for DNA-SWCNT sensors.

Analytical Approaches for Monitoring DNA-Protein Interactions.

DNA-protein interactions play a critical role in cellular regulation. We herein review existing analytical methods for investigating these interactions, highlighting methods such as chromatin immunoprecipitation and yeast-one-hybrid that are used to identify undiscovered DNA-protein interactions. We summarize the most common approaches for characterizing known interactions based on DNA-protein structure, thermodynamic and kinetic measurements, and dynamic binding assays. We discuss techniques in optical imaging as well as representative methods, such as eletrophoretic mobility shift assay and surface plasmon resonance. The advantages and disadvantages of these techniques are used to assess a proposed optical platform based on single-walled carbon nanotube (SWCNT) fluorescence.

Directed evolution of the optoelectronic properties of synthetic nanomaterials.

Directed evolution is a powerful approach to tailor protein properties toward new or enhanced functions. Herein, we use directed evolution to engineer the optoelectronic properties of DNA-wrapped single-walled carbon nanotube sensors through DNA mutation. This approach leads to an improvement in the fluorescence intensity of 56% following two evolution cycles.

Development of a novel thermal-sensitive multifunctional liposome with antibody conjugation to target EGFR-expressing tumors.

A novel EGFR-targeting, thermal-sensitive multifunctional liposome (TSML) was developed based on manganese-doped magnetism-engineered iron oxide nanoparticles (MnMEIOs) and gold nanorods (AuNRs) for efficient photothermal therapy and magnetic resonance (MR) imaging. An Erbitux-conjugated TSML (Erb-TSML) was encapsulated with doxorubicin and gold nanorods conjugated with manganese-doped magnetism-engineered iron oxide nanoparticles, for theranostic applications of EGFR-positive tumors. The Erb-TSML selectively targeted EGFR-positive tumors and promoted tumor destruction by laser activation. Using confocal microscopy, MR and optical imaging, we demonstrated that Erb-TSML specifically bound to A431 tumor cells. No signs of major morphological damages to the normal tissues were observed in mice treated with Erb-TSML and laser, indicating this theranostic platform protected heart against doxorubicin-induced toxicity to normal tissues. These results indicate that the Erb-TSML may be a promising diagnostic and therapeutic platform for EGFR-overexpressing tumors.

Restriction Enzyme Analysis of Double-Stranded DNA on Pristine Single-Walled Carbon Nanotubes.

Nanoprobes such as single-walled carbon nanotubes (SWCNTs) are capable of label-free detection that benefits from intrinsic and photostable near-infrared fluorescence. Despite the growing number of SWCNT-based applications, uncertainty surrounding the nature of double-stranded DNA (dsDNA) immobilization on pristine SWCNTs has limited their use as optical sensors for probing DNA-protein interactions. To address this limitation, we study enzyme activity on unmodified dsDNA strands immobilized on pristine SWCNTs. Restriction enzyme activity on various dsDNA sequences was used to verify the retention of the dsDNA's native conformation on the nanotube surface and to quantitatively compare the degree of dsDNA accessibility. We report a 2.8-fold enhancement in initial enzyme activity in the presence of surfactants. Förster resonance electron transfer (FRET) analysis attributes this enhancement to increased dsDNA displacement from the SWCNT surface. Furthermore, the accessibility of native dsDNA was found to vary with DNA configuration and the spacing between the restriction site and the nanotube surface, with a minimum spacing of four base pairs (bp) from the anchoring site needed to preserve enzyme activity. Molecular dynamics (MD) simulations verify that the anchored dsDNA remains within the vicinity of the SWCNT, revealing an unprecedented bimodal displacement of the bp nearest to SWCNT surface. Together, these findings illustrate the successful immobilization of native dsDNA on pristine SWCNTs, offering a new near-infrared platform for exploring vital DNA processes.

[Complementary Therapy for Patients With Limb Trauma Pain in the Emergency Department].

Acute pain is the main cause for patient visits to the emergency department. An important issue faced in emergency medical care is to resolve patient pain. Moreover, limb traumatic pain is noted in 58-66% of emergency trauma patients. This paper explored the effectiveness of using non-pharmacological complementary interventions, including body operation therapies that used trauma limb care and therapeutic touch and mind-body interventions, in order to strengthen the support system and create a healing environment. The experience described herein is hoped to facilitate the provision of multifaceted emergency care for patients with limb trauma pain in the emergency department in order to improve the comfort and return rate of patients.

Living on the Edge: Re-shaping the Interface of Synthetic Biology and Nanotechnology.

A new team of researchers at EPFL is taking an 'anti-disciplinary' approach to creating optical devices. These devices take advantage of the synergy in tuning both nano- and bio-material properties, coupling the advantages of two growing, albeit traditionally distinct, fields. With applications spanning from biosensing and microarray assays to living photovoltaics, the Laboratory of NanoBiotechnology (LNB) is uncovering an unexplored space for the next generation of chemical analytics and light-harvesting technologies.

Kinetically Enhanced Approach for Rapid and Tunable Self-Assembly of Rod-Coil Block Copolymers.

A facile approach is reported to process rod-coil block copolymers (BCPs) into highly ordered nanostructures in a rapid, low-energy process. By introducing a selective plasticizer into the rod-coil BCPs during annealing, both the annealing temperature and time to achieve thermodynamic equilibrium and highly ordered structures can be decreased. This process improvement is attributed to enhanced chain mobility, reduced rod-rod interaction, and decreased rod-coil interaction from the additive. The novel method is based on kinetically facilitating thermodynamic equilibrium. The process requires no modification of polymer structure, indicating that a wide variety of desired polymer functionalities can be designed into BCPs for specific applications.

Self-aggregation of amphiphilic [60]fullerenyl focal point functionalized PAMAM dendrons into pseudodendrimers: DNA binding involving dendriplex formation.

In this research, we successfully performed a "click" synthesis of amphiphilic poly(amido amine) dendron-bearing fullerenyl conjugate (C60 G1 ) using a copper(I)-catalyzed azide-alkyne cycloaddition reaction. The strong hydrophobicity of the C60 moiety induces self-assembly of C60 G1 into core-shell-like "pseudodendrimers" with a uniform size distribution and positively charged peripherals. The pseudodendrimers were well-characterized by atomic force microscopy (AFM), transmission electron microscopy, and dynamic light scattering. On the basis of electrostatic interactions, the polycationic C60 G1 assembly can serve as a nonviral gene vector. An ethidium bromide displacement assay and agarose gel electrophoresis both indicated that C60 G1 assembly forms stable complexes with the cyclic reporter gene (pEGFP-C1) at low nitrogen-to-phosphorous (N/P) ratios. AFM analysis revealed a dynamic complex-formation process, and confirmed the synthesis of C60 G1 /pEGFP-C1 hybrids with a particle dimensions less than 200 nm. Fluorescence microscopy and flow cytometry revealed that 51% of HeLa and 43% of MCF-7 cells are positive to the YOYO-1-labeled hybrids at an N/P ratio of 2, being comparable to TurboFect-mediated delivery.