Biporous silica nanostructure-induced nanovortex in microfluidics for nucleic acid enrichment, isolation, and PCR-free detection.

Efficient pathogen enrichment and nucleic acid isolation are critical for accurate and sensitive diagnosis of infectious diseases, especially those with low pathogen levels. Our study introduces a biporous silica nanofilms-embedded sample preparation chip for pathogen and nucleic acid enrichment/isolation. This chip features unique biporous nanostructures comprising large and small pore layers. Computational simulations confirm that these nanostructures enhance the surface area and promote the formation of nanovortex, resulting in improved capture efficiency. Notably, the chip demonstrates a 100-fold lower limit of detection compared to conventional methods used for nucleic acid detection. Clinical validations using patient samples corroborate the superior sensitivity of the chip when combined with the luminescence resonance energy transfer assay. The enhanced sample preparation efficiency of the chip, along with the facile and straightforward synthesis of the biporous nanostructures, offers a promising solution for polymer chain reaction-free detection of nucleic acids.

Toxicity and Biodistribution of Fragmented Polypropylene Microplastics in ICR Mice.

Currently, polypropylene (PP) is used in various products, thus leading to high daily exposure in humans. Thus, it is necessary to evaluate the toxicological effects, biodistribution, and accumulation of PP microplastics in the human body. In this study, administration of two particle sizes of PP microplastics (approximately 5 and 10-50 µm) did not lead to any significant changes in several toxicological evaluation parameters, including body weight and pathological examination, compared with the control group in ICR mice. Therefore, the approximate lethal dose and no-observed-adverse-effect level of PP microplastics in ICR mice were established as ≥2000 mg/kg. Furthermore, we manufactured cyanine 5.5 carboxylic acid (Cy5.5-COOH)-labeled fragmented PP microplastics to monitor real-time in vivo biodistribution. After oral administration of the Cy5.5-COOH-labeled microplastics to the mice, most of the PP microplastics were detected in the gastrointestinal tract and observed to be out of the body after 24 h in IVIS Spectrum CT. Therefore, this study provides a new insight into the short-term toxicity, distribution, and accumulation of PP microplastics in mammals.

On-site bioaerosol sampling and detection in microfluidic platforms.

As the recent coronavirus disease (COVID-19) pandemic and several severe illnesses such as Middle East respiratory syndrome coronavirus (MERS-CoV), Influenza A virus (IAV) flu, and severe acute respiratory syndrome (SARS) have been found to be airborne, the importance of monitoring bioaerosols for the control and prevention of airborne epidemic diseases outbreaks is increasing. However, current aerosol collection and detection technologies may be limited to on-field use for real-time monitoring because of the relatively low concentrations of targeted bioaerosols in air samples. Microfluidic devices have been used as lab-on-a-chip platforms and exhibit outstanding capabilities in airborne particulate collection, sample processing, and target molecule analysis, thereby highlighting their potential for on-site bioaerosol monitoring. This review discusses the measurement of airborne microorganisms from air samples, including sources and transmission of bioaerosols, sampling strategies, and analytical methodologies. Recent advancements in microfluidic platforms have focused on bioaerosol sample preparation strategies, such as sorting, concentrating, and extracting, as well as rapid and field-deployable detection methods for analytes on microfluidic chips. Furthermore, we discuss an integrated platform for on-site bioaerosol analyses. We believe that our review significantly contributes to the literature as it assists in bridging the knowledge gaps in bioaerosol monitoring using microfluidic platforms.

Flexible 3D Nanonetworked Silica Film as a Polymer-Free Drug-Eluting Stent Platform to Effectively Suppress Tissue Hyperplasia in Rat Esophagus.

Loading and eluting drugs on self-expandable metallic stents (SEMSs) can be challenging in terms of fabrication, mechanical stability, and therapeutic effects. In this study, a flexible 3D nanonetworked silica film (NSF) capable of withstanding mechanical stress during dynamic expansion is constructed to function as a drug delivery platform on an entire SEMS surface. Despite covering a broad curved area, the synthesized NSF is defect-free and thin enough to increase the stent strut diameter (110 µm) by only 0.4 percent (110.45 µm). The hydrophobic modification of the surface enables loading of 4.7 times the sirolimus (SRL) concentration in NSF than Cypher, polymer-coated commercial stent, which is based on the same thickness of coating layer. Furthermore, SRL-loaded NSF exhibits a twofold delay in release compared to the control group without NSF. The SRL-loaded NSF SEMS significantly suppresses stent-induced tissue hyperplasia than the control SEMS in the rat esophagus (all variables, p < 0.05). Thus, the developed NSF is a promising polymer-free drug delivery platform to efficiently treat esophageal stricture.

Correspondence between Parents' and Adolescents' Sleep Duration.

This study explored the correspondence between adolescents' sleep duration and that of their parents and identified the factors affecting the appropriate sleep duration for adolescents. The data of 795 adolescents from the Korea National Health and Nutrition Examination Survey (2015-2018) were analyzed. We used Cohen's kappa coefficient to measure the correspondence between adolescents' sleep duration and that of their parents. A multiple logistic regression analysis was used to identify the factors affecting adequate sleep duration among adolescents. Our study found that factors such as adolescents' gender, father's education level, and drinking among adolescents and parents influenced the adolescents' sleep duration. Second, a higher correspondence between the sleep duration of adolescents and that of mothers (Kappa = 0.213, p < 0.001) was found compared to that of fathers (Kappa = 0.064, p = 0.031). Finally, an adequate sleep duration among adolescents' mothers was a major factor that influenced the adequate sleep duration of adolescents (OR = 2.494, 95% CI = 1.850-3.362, p < 0.001). Therefore, when organizing adolescent sleep education and management programs in various community sleep management institutions, the main caregiver's sleep duration management and family drinking management should be combined. Additionally, gender equality awareness should be improved for parenting, including monitoring adolescents' sleep accordingly.

Macroscopic Assembly of Sericin toward Self-Healable Silk.

Silk contains an adhesive glycoprotein, silk sericin, in which silk fibroins can be enfolded and chemically stabilized. Silk sericin is gaining importance as the material for the creation of functional bioscaffolds. However, the assembly of silk sericin is generally limited to the blend of polymers or proteins due to its inherent poor mechanical strength. Here, we report a simple macroscopic controlled assembly of silk sericin fibers based on their secondary structure via wet-spinning. In addition, plasticization of silk sericin using glycerol immobilized with glutaraldehyde was found to induce dimensional stability, affording stable linear fibers with self-adhesion. Furthermore, cyclo-phenylalanine nanowires were incorporated into the silk sericin dope for a practical demonstration of their potential in artificial silk production with superstructure formation. The physicochemical characteristics of the spun fibers have also been elucidated using Fourier-transform infrared spectroscopy, electron microscopy, tensile test, differential scanning calorimetry, and 2D X-ray diffraction.

Genetic heterogeneity in Leigh syndrome: Highlighting treatable and novel genetic causes.

Leigh syndrome (LS), the most common childhood mitochondrial disorder, has characteristic clinical and neuroradiologic features. Mutations in more than 75 genes have been identified in both the mitochondrial and nuclear genome, implicating a high degree of genetic heterogeneity in LS. To profile these genetic signatures and understand the pathophysiology of LS, we recruited 64 patients from 62 families who were clinically diagnosed with LS at Seoul National University Children's Hospital. Mitochondrial genetic analysis followed by whole-exome sequencing was performed on 61 patients. Pathogenic variants in mitochondrial DNA were identified in 18 families and nuclear DNA mutations in 22. The following 17 genes analyzed in 40 families were found to have genetic complexity: MTATP6, MTND1, MTND3, MTND5, MTND6, MTTK, NDUFS1, NDUFV1, NDUFAF6, SURF1, SLC19A3, ECHS1, PNPT1, IARS2, NARS2, VPS13D, and NAXE. Two treatable cases had biotin-thiamine responsive basal ganglia disease, and another three were identified as having defects in the newly recognized genes (VPS13D or NAXE). Variants in the nuclear genes that encoded mitochondrial aminoacyl tRNA synthetases were present in 27.3% of cases. Our findings expand the genetic and clinical spectrum of LS, showing genetic heterogeneity and highlighting treatable cases and those with novel genetic causes.

Indoxyl Sulfate-Induced Extracellular Vesicles Released from Endothelial Cells Stimulate Vascular Smooth Muscle Cell Proliferation by Inducing Transforming Growth Factor-Beta Production.

Vascular access stenosis predominantly occurs as a result of neointimal hyperplasia (NH) formation at the anastomosis. Moreover, in the presence of NH, transforming growth factor-beta (TGF-ß) promotes vascular smooth muscle cell (VSMC) proliferation. Extracellular vesicles (EVs) released by endothelial cells are closely associated with vascular dysfunction. Here, we investigated the effects of EVs on TGF-ß signaling and VSMC proliferation. Specifically, EVs were collected from the culture medium of indoxyl sulfate (IS)-treated human umbilical vein endothelial cells and used (2 × 106) to stimulate human aortic smooth muscle cells (SMCs) (1 × 106). Western blotting was performed to assess the levels of Akt, ERK1/2, p38 MAPK, and Smad3. BrdU proliferation assays, quantitative PCR, and ELISA assays were performed to evaluate SMC proliferation and TGF-ß production. The IS-induced EVs stimulated the proliferation of aortic SMCs in a concentration-dependent manner. The EVs both contained TGF-ß and promoted TGF-ß production by SMCs by phosphorylating Akt, ERK1/2, p38 MAPK, and Smad3, which was significantly inhibited by an anti-TGF-ß antibody. SMC proliferation was suppressed by both an anti-TGF-ß antibody and inhibitors of the downstream factors. These results suggest that EVs are involved in the pathogenesis of vascular access stenosis by modulating TGF-ß signaling in VSMCs under uremic conditions.

Korean National Oral Health Survey Data on the Symmetry of Primary Dentition Surface Caries.

OBJECTIVES: This study evaluated the intraoral symmetry of dental caries in primary teeth as part of a study of caries patterns in primary dentition. STUDY DESIGN: The data for 4,800 5-year-old and 4,379 8-year-old children in this study were from the 2012 Korean national oral health survey. Pearson correlation coefficients of the decayed and filled surface (dfs) values ranged from 0.436 (lower primary canines) to 0.835 (upper primary central incisors) for the right and left primary teeth and from 0.084 (right primary central incisor) to 0.457 (left primary second molar) for the upper and lower primary dentition (P< 0.01). RESULTS: The upper and lower dfs values differed significantly (P< 0.05) when the right and left primary second molars were excluded. The left or right primary data without caries ranged from 56.4% (lower of first and second primary molars) to 99.7% (lower primary central incisors). The bilateral caries among cases with one or more in the right or left primary teeth ranged from 25.0% (lower lateral primary incisor) to 72.7% (upper primary central incisors). CONCLUSIONS: These results suggested that dental caries in primary teeth show bilateral symmetry and differences in the degree of symmetry according to the teeth set or surface set of the homologous teeth.

Epidermal growth factor-mediated Rab25 pathway regulates integrin ß1 trafficking in colon cancer.

BACKGROUND: Integrins play a critical role in carcinogenesis. Integrin ß1 localization is regulated by the guanosine-5'-triphosphate hydrolase Rab25 and integrin ß1 levels are elevated in the serum of colon cancer patients; thus, the present study examined the effects of epidermal growth factor (EGF) and Rab25 on integrin ß1 localization in colon cancer cells. METHODS: HCT116 human colon cancer cells were treated with increasing concentrations of EGF, and cell proliferation and protein expression were monitored by MTT and western blot analyses, respectively. Cell fractionation was performed to determine integrin ß1 localization in the membrane and cytosol. Integrin ß1 extracellular shedding was monitored by enzyme-linked immunosorbent assays (ELISAs) with culture supernatants from stimulated cells. HCT116 cells were transfected with Rab25-specific siRNA to determine the significance of Rab25 in integrin ß1 trafficking in the presence of EGF. RESULTS: Total integrin ß1 expression increased in response to EGF and subsequently decreased at 24 h post-stimulation. A similar decrease was observed in purified membrane fractions, whereas no changes were observed in cytosolic levels. ELISAs using media from stimulated cell cultures demonstrated increased integrin ß1 levels corresponding to the decrease observed in membrane fractions, suggesting that EGF induces integrin receptor shedding. EGF stimulation in Rab25-knockdown cells resulted in integrin ß1 accumulation in the membrane, suggesting that Rab25 promotes integrin endocytosis. CONCLUSIONS: Integrin ß1 is shed from colon cancer cells in response to EGF stimulation in a Rab25-dependent manner. These results further the present understanding of the role of integrin ß1 in colon cancer progression.

Performance standard-based validation study for local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method.

Local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method (LLNA: BrdU-FCM) is a modified non-radioisotopic technique with the additional advantages of accommodating multiple endpoints with the introduction of FCM, and refinement and reduction of animal use by using a sophisticated prescreening scheme. Reliability and accuracy of the LLNA: BrdU-FCM was determined according to OECD Test Guideline (TG) No. 429 (Skin Sensitization: Local Lymph Node Assay) performance standards (PS), with the participation of four laboratories. Transferability was demonstrated through successfully producing stimulation index (SI) values for 25% hexyl cinnamic aldehyde (HCA) consistently greater than 3, a predetermined threshold, by all participating laboratories. Within- and between-laboratory reproducibility was shown using HCA and 2,4-dinitrochlorobenzene, in which EC2.7 values (the estimated concentrations eliciting an SI of 2.7, the threshold for LLNA: BrdU-FCM) fell consistently within the acceptance ranges, 0.025-0.1% and 5-20%, respectively. Predictive capacity was tested using the final protocol version 1.3 for the 18 reference chemicals listed in OECD TG 429, of which results showed 84.6% sensitivity, 100% specificity, and 88.9% accuracy compared with the original LLNA. The data presented are considered to meet the performance criteria for the PS, and its predictive capacity was also sufficiently validated.

Predictive capacity of a non-radioisotopic local lymph node assay using flow cytometry, LLNA:BrdU-FCM: Comparison of a cutoff approach and inferential statistics.

In order for a novel test method to be applied for regulatory purposes, its reliability and relevance, i.e., reproducibility and predictive capacity, must be demonstrated. Here, we examine the predictive capacity of a novel non-radioisotopic local lymph node assay, LLNA:BrdU-FCM (5-bromo-2'-deoxyuridine-flow cytometry), with a cutoff approach and inferential statistics as a prediction model. 22 reference substances in OECD TG429 were tested with a concurrent positive control, hexylcinnamaldehyde 25%(PC), and the stimulation index (SI) representing the fold increase in lymph node cells over the vehicle control was obtained. The optimal cutoff SI (2.7≤cutoff <3.5), with respect to predictive capacity, was obtained by a receiver operating characteristic curve, which produced 90.9% accuracy for the 22 substances. To address the inter-test variability in responsiveness, SI values standardized with PC were employed to obtain the optimal percentage cutoff (42.6≤cutoff <57.3% of PC), which produced 86.4% accuracy. A test substance may be diagnosed as a sensitizer if a statistically significant increase in SI is elicited. The parametric one-sided t-test and non-parametric Wilcoxon rank-sum test produced 77.3% accuracy. Similarly, a test substance could be defined as a sensitizer if the SI means of the vehicle control, and of the low, middle, and high concentrations were statistically significantly different, which was tested using ANOVA or Kruskal-Wallis, with post hoc analysis, Dunnett, or DSCF (Dwass-Steel-Critchlow-Fligner), respectively, depending on the equal variance test, producing 81.8% accuracy. The absolute SI-based cutoff approach produced the best predictive capacity, however the discordant decisions between prediction models need to be examined further.

Dental fear & anxiety and dental pain in children and adolescents; a systemic review.

BACKGROUND: There are few previous studies investigating the relationship of dental fear and anxiety (DFA) with dental pain among children and adolescents. To address this issue, we examined the literature published between November 1873 and May 2015 to evaluate the prevalence of DFA and dental pain among children and adolescents, and their relationships with age and sex. METHODS: We performed a broad search of the PubMed database using 3 combinations of the search terms dental fear, anxiety, and dental pain and prevalence. A large proportion of the identified articles could not be used for the review due to inadequate end points or measures, or because of poor study design. Thirty-two papers of acceptable quality were identified and reviewed. RESULTS: We found that the prevalence of DFA was estimated to be 10%, with a decrease in prevalence with age. It was more frequently seen in girls, and was related to dental pain. CONCLUSIONS: We concluded that dental fear, anxiety, and pain are common, and several psychological factors are associated with their development. In order to better understand these relationships, further clinical evaluations and studies are required.